TW202235069A - A composition containing lactobacillus spp. and its use for preventing and/or improving anti-aging of skin - Google Patents
A composition containing lactobacillus spp. and its use for preventing and/or improving anti-aging of skin Download PDFInfo
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本發明關於包含乳酸桿菌的組成物及預防及/或改善皮膚老化的用途；更明確地說，係關於一種包含胚芽乳酸桿菌（ Lactobacillus plantarum）、乳雙歧桿菌（ Bifidobacterium lactis）或其組合的組成物，以用於預防及/或改善隨著老化增加的糖化壓力所造成的皮膚上的皺紋、細紋、斑或鬆弛等。 The present invention relates to a composition comprising Lactobacillus and its use for preventing and/or improving skin aging; more specifically, it relates to a composition comprising Lactobacillus plantarum , Bifidobacterium lactis or a combination thereof It is used to prevent and/or improve wrinkles, fine lines, spots or sagging on the skin caused by the glycation stress that increases with aging.
糖化作用的機制Mechanism of Glycation
常見的糖化反應可分為三個階段：（1）第一階段為糖類分子與帶有胺基團物質交互作用，而產生阿馬道里 （Amadori） 產物的過程；（2）第二階段為 Amadori 產物經過降解，而產生高度活性之羰基化合物，例如乙二醛 （GO）、甲基乙二醛 （MG） 等之過程；（3）第三階段為糖化中期的高度活性之羰基化合物與蛋白質或胺基酸交互作用反應生成穩定物質之過程，其中與蛋白質反應生成之物質稱為糖化蛋白、與胺基酸反應或糖化蛋白之糖化殘基 （residues） 裂解生成之小分子物質稱為糖化分子。在糖分子與糖代謝過程中產生之活性羰基物質與蛋白質等具胺基之分子交互反應後之產物，即高度糖化終產物 （advanced glycation endproducts, AGEs）。糖化反應及其產物的生成路徑與機制顯示如圖1。Common saccharification reactions can be divided into three stages: (1) The first stage is the process in which carbohydrate molecules interact with substances with amine groups to produce Amadori products; (2) The second stage is the process of Amadori The product is degraded to produce highly active carbonyl compounds, such as glyoxal (GO), methylglyoxal (MG), etc.; (3) The third stage is the highly active carbonyl compound and protein or The process of amino acid interaction reaction to generate stable substances, in which the substance formed by reacting with protein is called glycated protein, and the small molecular substance formed by reaction with amino acid or cleavage of glycated residues (residues) of glycated protein is called glycated molecule. The products of the interaction between sugar molecules and active carbonyl substances produced in the process of sugar metabolism and molecules with amino groups such as proteins, are advanced glycation end products (AGEs). The formation pathway and mechanism of the saccharification reaction and its products are shown in Figure 1.
AGEs 對生理之影響則稱為糖化壓力 （glycative stress）。 AGEs 對生理的影響包含：（1）具有促進蛋白質交聯之特性。AGEs可作為蛋白質中不同胜肽鏈之架橋分子，促使聚集反應 （protein aggregation） 或組織剛性 （tissue stiffness），使蛋白質失去原有之功能特性。（2） 蛋白結合 （Protein-bound）的 AGEs 可透過活化細胞膜上之高度糖化中產物受器 （the receptor for AGEs, RAGE）、oligosaccharyl transferase complex protein 48 （OST-48, generally known as AGER1）、80 K-H protein （AGER2）、galectin-3 （AGER3） 等受器或直接促進 ROS 生成，影響細胞生理。由於 RAGE 活化與許多疾病具有正相關性，因此， AGEs 與 RAGE 交互作用及其於疾病中扮演重要角色。The physiological effects of AGEs are called glycation stress. The physiological effects of AGEs include: (1) It has the property of promoting protein cross-linking. AGEs can act as bridge molecules between different peptide chains in proteins to promote protein aggregation or tissue stiffness, causing proteins to lose their original functional properties. (2) Protein-bound AGEs can activate the receptor for AGEs (RAGE), oligosaccharyl transferase complex protein 48 (OST-48, generally known as AGER1), 80 K-H protein (AGER2), galectin-3 (AGER3) and other receptors may directly promote ROS production and affect cell physiology. Since RAGE activation is positively correlated with many diseases, AGEs interact with RAGE and play an important role in diseases.
糖化作用與皮膚老化Glycation and Skin Aging
特別針對上述（1）之 AGEs 生理作用，常被研究於糖尿病與慢性腎臟病併發之心血管疾病、老化、或皮膚病以及神經退化等相關疾病中。許多研究證實糖化壓力與多種疾病進程具相關性，相關疾病包含糖尿病、肥胖；神經與認知疾病、心血管疾病、腎病變、肝病變、部分癌症 （乳癌、大腸癌、胰臟癌等）。許多研究證實糖化壓力亦與生物體的老化具高度關聯性。在內部生理反應中，包括動脈粥樣硬化、白內障或其他相關炎症等。在外部表徵上，糖化壓力則會使人體的皮膚肌理粗糙化或鬆弛、膚色黯淡且長斑點、唇周乾燥與脫皮等。這是因為，AGEs沈澱附著於皮膚組織中，而與被稱為RAGE之受體鍵結，在皮膚或其他組織中引起炎症性變化。同時在生成AGEs之過程中，會於膠原蛋白之分子內及分子間形成無秩序的交聯，對膠原蛋白引發物理性、生理性變化（改性）。即造成皺紋、肌膚粗糙、暗沈、或緊緻性降低等肌膚的老化。又，藉由糖化反應，在分子內及分子間形成有無秩序交聯的膠原蛋白，除了會失去彈力性外，同時會因生命體內的蛋白分解酶而變得難以被分解，其結果恐會導致肌膚轉換速度的延遲。因此，在醫藥領域中基於研究糖化反應與老化的關聯，嘗試藉由抑制糖化反應來預防皮膚老化，來開發另一種替代性的治療途徑。Especially for the physiological effects of AGEs in (1) above, it is often studied in related diseases such as cardiovascular disease, aging, skin disease and neurodegeneration complicated by diabetes and chronic kidney disease. Many studies have confirmed that glycation stress is associated with various disease processes, including diabetes, obesity; neurological and cognitive diseases, cardiovascular diseases, kidney diseases, liver diseases, and some cancers (breast cancer, colorectal cancer, pancreatic cancer, etc.). Many studies have confirmed that glycation stress is also highly correlated with the aging of organisms. In the internal physiological response, including atherosclerosis, cataract or other related inflammation, etc. In terms of external manifestations, glycation stress will cause the human body's skin texture to become rough or loose, dark complexion with spots, dryness and peeling around the lips, etc. This is because AGEs deposit and attach to skin tissue, and bind to receptors called RAGE, causing inflammatory changes in the skin or other tissues. At the same time, in the process of generating AGEs, disordered cross-links will be formed within and between molecules of collagen, causing physical and physiological changes (modifications) to collagen. That is, it causes skin aging such as wrinkles, rough skin, dullness, or loss of firmness. In addition, due to the saccharification reaction, the collagen with disordered cross-links formed in the molecule and between the molecules will not only lose its elasticity, but also become difficult to be decomposed by the proteolytic enzymes in the living body, and the result may lead to Delay in skin transition speed. Therefore, in the field of medicine, based on the research on the relationship between glycation reaction and aging, it is attempted to prevent skin aging by inhibiting glycation reaction to develop another alternative treatment approach.
現有的抗糖化原料Existing anti-glycation raw materials
目前市場上被研究具抗糖化功效之原料，分為化學性合成結構或植物來源之天然物。植物來源例如肉桂 （ Cinnamomum cassia）、小茴香 （ Cuminum cyminum）、迷迭香 （Rosemary）、魚腥草（ Houttuynia cordata）等，萃取出來的天然物舉例如薑黃素 （curcumin）、芸香苷 （Rutin）、槲皮苷（quercitrin）。 Raw materials currently on the market that are researched to have anti-glycation effects are divided into chemically synthesized structures or natural products derived from plants. Plant sources such as cinnamon ( Cinnamomum cassia ), cumin ( Cuminum cyminum ), rosemary (Rosemary), Houttuynia cordata ( Houttuynia cordata ), etc., extracted natural products such as curcumin (curcumin), rutin (Rutin) , quercitrin (quercitrin).
對於化學合成的抗糖化產品，消費者普遍接受度不高，且依然有健康上的疑慮。但是，植物性來源的抗糖化天然物，卻又往往因為使用大量的溶劑萃取，而難以避免會有溶劑殘留的疑慮。又或者，要使溶劑低劑量殘留的方法與設備門檻極高，造成產品的價格也偏高。因此，不論是化學合成或天然萃取的產品，皆無法充分滿足市場大眾的需求。Consumers generally do not accept chemically synthesized anti-glycation products, and there are still health concerns. However, anti-glycation natural products derived from plants are often extracted with a large amount of solvents, so it is difficult to avoid the doubts of solvent residues. Or, the method and equipment threshold for making low-dose solvent residues is extremely high, resulting in high product prices. Therefore, neither chemically synthesized nor naturally extracted products can fully meet the needs of the market public.
況且，由於糖化壓力與皮膚老化、粗糙化、鬆弛、脫皮等困擾及其他許多疾病之發展具相關性，因此開發抗糖化之原料非常具有市場價值。且市場上也很期待以微生物來源之抗糖化功能組合物作為新穎材料，來擴充市場上對於抗糖化產品選擇的多樣性。Moreover, since glycation stress is related to skin aging, roughness, sagging, peeling and many other diseases, the development of anti-glycation raw materials is of great market value. And the market is also looking forward to using the anti-glycation functional composition derived from microorganisms as a novel material to expand the diversity of anti-glycation product choices on the market.
乳酸桿菌屬Lactobacillus
乳酸桿菌屬（ Lactobacillus spp.） 存在於一般環境中，其能將碳水化合物發酵成乳酸，因此多應用於發酵食品的製造。早在1900年代，就已發現此菌對於人體健康具有多重益處，具有幫助消化、改善腸道健康的功效，而被提倡多食用乳酸桿菌以改善消化環境、促進腸道蠕動。經研究後又發現，由於乳酸桿菌能分解醣類（舉例如乳糖、葡萄糖、蔗糖、果糖），因此可產生乳酸和醋酸，並藉此酸化腸道的環境，而達到抑制害菌於腸道中增殖，以及調整腸內菌叢平衡的有益效果。 Lactobacillus spp. exists in the general environment and can ferment carbohydrates into lactic acid, so it is mostly used in the manufacture of fermented foods. As early as the 1900s, it was discovered that this bacterium has multiple benefits for human health, such as helping digestion and improving intestinal health, and it is advocated to eat more Lactobacillus to improve the digestive environment and promote intestinal peristalsis. After research, it was found that because Lactobacillus can decompose sugars (such as lactose, glucose, sucrose, fructose), it can produce lactic acid and acetic acid, and thereby acidify the intestinal environment, thereby inhibiting the proliferation of harmful bacteria in the intestinal tract , and the beneficial effect of adjusting the balance of intestinal flora.
縱使同為乳酸桿菌屬，若為不同種、或甚至是同種但不同菌株都彼此具有不一樣的特徵，對人體亦會有不同的影響。例如植物乳桿菌（ Lactobacillus plantarum），有文獻指出 L. plantarum PH04具降低血中膽固醇能力；在一另篇文獻證實 L.plantarum 299V可減輕IL-10缺乏型小鼠之結腸炎症狀；又在別的文獻指出 L.plantarum 10hk2可提高前發炎介質，如IL-1β、IL-6和TNF-α，及抗發炎介白素-10（IL-10），達到抗發炎效果；甚至還有文獻證實 L. plantarum K21具有降低血中膽固醇、三酸甘油脂含量，以及具有抗發炎之功效。上述列舉的多種特定菌株都屬植物乳桿菌而為同種，但卻因其中的有效成分不同得以應用於不同的疾病或病症。 Even if they belong to the same genus of Lactobacillus, if they are of different species, or even of the same species but different strains have different characteristics from each other, they will have different effects on the human body. For example, Lactobacillus plantarum ( Lactobacillus plantarum ), some literature pointed out that L. plantarum PH04 has the ability to lower blood cholesterol; another literature confirmed that L. plantarum 299V can reduce the symptoms of colitis in IL-10-deficient mice; The literature pointed out that L.plantarum 10hk2 can increase pro-inflammatory mediators, such as IL-1β, IL-6 and TNF-α, and anti-inflammatory interleukin-10 (IL-10), to achieve anti-inflammatory effects; there are even literatures confirmed L. plantarum K21 can lower blood cholesterol and triglyceride levels, and has anti-inflammatory effects. The various specific strains listed above belong to the same species of Lactobacillus plantarum, but they can be applied to different diseases or diseases because of the different active ingredients therein.
植物乳桿菌Lactobacillus plantarum
植物乳桿菌存在於唾液及許多發酵食品中。植物乳桿菌有很強的適應性，一種革蘭氏陽性厭氧菌，可以在攝氏10至45度的溫度下生長，並且適應3.2-4.2或更高的pH值水平。植物乳桿菌首次自唾液被分離，隨後的研究發現此細菌的部分特定菌株可作為益生菌以應用於抗氧化、保持腸道通透性、抑制腸道內產氣細菌的生長，還能增加海馬體的腦源性神經營養因子，以應用在抑鬱症的治療。Lactobacillus plantarum is found in saliva and many fermented foods. Lactobacillus plantarum is highly adaptable, a Gram-positive anaerobic bacterium that can grow at temperatures ranging from 10 to 45 degrees Celsius and is adapted to pH levels of 3.2-4.2 or higher. Lactobacillus plantarum was isolated from saliva for the first time, and subsequent studies found that some specific strains of this bacterium can be used as probiotics for anti-oxidation, maintaining intestinal permeability, inhibiting the growth of gas-producing bacteria in the intestinal tract, and increasing hippocampal The body's brain-derived neurotrophic factor can be applied in the treatment of depression.
雙歧桿菌屬Bifidobacterium
雙歧桿菌屬（ Bifidobacterium）廣泛存在於人和動物的消化道、***和口腔等環境中。其為革蘭氏陽性、不運動、細胞呈杆狀、一端有時呈分叉狀且嚴格厭氧。此細菌早在1899年就從健康應而的糞便中分離出來，隨後的研究發現此細菌的部分特定菌株可作為益生菌以應用於食品、醫藥和飼料等領域。 Bifidobacterium ( Bifidobacterium ) widely exists in the digestive tract, vagina and oral cavity of humans and animals. It is Gram-positive, immobile, has rod-shaped cells, sometimes forked at one end, and is strictly anaerobic. This bacterium was isolated from the feces of healthy people as early as 1899, and subsequent research found that some specific strains of this bacterium can be used as probiotics in the fields of food, medicine and feed.
為改善隨著年齡增長而出現的皮膚問題，例如皺紋、細紋、斑、鬆弛等，以及開發新材料以突破現有材料種類的侷限性，本發明提供一種預防及/或改善皮膚老化的組成物，其包含胚芽乳酸桿菌（ Lactobacillus plantarum）、乳雙歧桿菌（ Bifidobacterium lactis）或其組合。 In order to improve skin problems that occur with age, such as wrinkles, fine lines, spots, sagging, etc., and to develop new materials to break through the limitations of existing materials, the present invention provides a composition for preventing and/or improving skin aging , comprising Lactobacillus plantarum , Bifidobacterium lactis or a combination thereof.
較佳地，該胚芽乳酸桿菌係菌株GKM3以寄存編號BCRC-910787寄存於財團法人食品工業發展研究所；該乳雙歧桿菌係菌株GKK2以寄存編號BCRC910826寄存於財團法人食品工業發展研究所。Preferably, the Lactobacillus plantarum strain GKM3 is deposited at the Food Industry Development Research Institute of the Foundation with the registration number BCRC-910787; the Bifidobacterium lactis strain GKK2 is deposited at the Food Industry Development Research Institute of the Foundation with the registration number BCRC910826.
較佳地，該組成物包含培養及/或發酵該乳酸桿菌的培養基成分Preferably, the composition comprises medium components for cultivating and/or fermenting the Lactobacillus
較佳地，該組成物進一步包含具有改善肌膚轉換作用及/或抑制糖化反應作用的植物萃取物或化合物。Preferably, the composition further includes plant extracts or compounds that have the effect of improving skin transformation and/or inhibiting glycation reaction.
較佳地，該具有改善肌膚轉換作用及/或抑制糖化反應作用的植物萃取物包含柏科、銀杏科、柑橘科、紫蘇科、魚腥草科、杜仲科的植物的萃取物。Preferably, the plant extract having the effect of improving skin transformation and/or inhibiting saccharification reaction includes plant extracts of Cupressaceae, Ginkgoceae, Citrus, Perillaceae, Houttuyniaceae, and Eucommiaceae.
較佳地，該柏科的植物包含羅漢柏；該銀杏科的植物包含銀杏；該柑橘科的植物包含黃檗、橙、葡萄柚、柚子及檸檬；該紫蘇科的植物包含薄荷、胡椒薄荷、紫蘇及迷迭香；該魚腥草科的植物包含魚腥草；該杜仲科的植物包含杜仲。Preferably, the plants of the family Cupressaceae include Podocarpus; the plants of the Ginkgo family include Ginkgo biloba; the plants of the Citrus family include corkberry, orange, grapefruit, pomelo and lemon; the plants of the Perilla family include mint, peppermint, perilla and rosemary; the plant of Houttuyniaceae comprises Houttuynia cordata; the plant of Eucommiaceae comprises Eucommia.
較佳地，該具有改善肌膚轉換作用及/或抑制糖化反應作用的化合物包含雌馬酚。Preferably, the compound having the effect of improving skin transformation and/or inhibiting glycation reaction includes equol.
較佳地，每單位的組成物中含有0.1重量%以上之乳酸桿菌，與合計為0.001重量%以上之具有改善肌膚轉換作用及/或抑制糖化反應作用的植物萃取物或化合物。Preferably, each unit of the composition contains more than 0.1% by weight of lactobacillus, and a total of more than 0.001% by weight of plant extracts or compounds that can improve skin transformation and/or inhibit glycation reaction.
較佳地，如前述之組成物係為醫藥組成物、化妝品、保養品、營養品或食品。Preferably, the aforementioned composition is a pharmaceutical composition, cosmetic, skin care product, nutritional product or food.
較佳地，該化妝品包含化妝水、洗面劑、美容液、乳液或乳霜。Preferably, the cosmetics include lotion, facial cleanser, beauty lotion, lotion or cream.
為使含有乳酸桿菌之組成物更容易施予有需要的受施者，本發明提供一種如前述之包含乳酸桿菌之組合物用於預防及/或改善皮膚老化的用途。In order to make it easier to administer the composition containing lactobacilli to recipients in need, the present invention provides a use of the aforementioned composition containing lactobacilli for preventing and/or improving skin aging.
較佳地，該包含乳酸桿菌之組成物以下列方法製備： （a）取一乳酸桿菌的菌落接種於培養基進行固態培養，而獲得菌體； （b）將步驟（a）的菌體接種於液體培養基進行液態培養，而獲得液態培養基與菌體的混合物；及 （c）將步驟（b）的液態培養基與菌體的混合物進行發酵培養，而獲得發酵全液。 Preferably, the composition comprising Lactobacillus is prepared by the following method: (a) Inoculate a colony of Lactobacillus in the culture medium for solid-state culture to obtain bacteria; (b) inoculating the bacteria in step (a) into a liquid medium for liquid culture to obtain a mixture of liquid medium and bacteria; and (c) Fermenting the mixture of the liquid medium and the cells in step (b) to obtain a complete fermentation solution.
較佳地，該方法進一步包含下列步驟： （d）將步驟（c）的發酵全液進行離心，而獲得菌泥及/或上清液；及 （e）將步驟（d）所得的該菌泥或該上清液進行冷凍乾燥，而獲得凍乾粉。 Preferably, the method further comprises the following steps: (d) centrifuging the whole fermentation liquid in step (c) to obtain the sludge and/or supernatant; and (e) Freeze-drying the sludge or the supernatant obtained in step (d) to obtain freeze-dried powder.
較佳地，該改善皮膚老化係藉由施予該包含乳酸桿菌之組成物來抑制或減緩受施者的糖化反應。Preferably, the skin aging is improved by administering the composition comprising lactobacilli to inhibit or slow down the glycation reaction of the recipient.
較佳地，該抑制或減緩受施者的糖化反應係指受施者的糖化前期產物的含量降低。Preferably, the inhibiting or slowing down of the subject's glycation response refers to the reduction of the content of pre-glycation products in the subject.
較佳地，該糖化前期產物包含阿馬道里 （Amadori）。Preferably, the pre-saccharification product comprises Amadori.
較佳地，該抑制或減緩受施者的糖化反應係指受施者的糖化中期雙羰基化合物的含量降低。Preferably, the inhibiting or slowing down of the glycation reaction of the recipient refers to the reduction of the content of dicarbonyl compounds in the mid-glycation stage of the recipient.
較佳地，該糖化中期雙羰基化合物包含乙二醛 （GO）、甲基乙二醛 （MG）或其組合。Preferably, the mid-saccharification dicarbonyl compound comprises glyoxal (GO), methylglyoxal (MG) or a combination thereof.
較佳地，該抑制或減緩受施者的糖化反應係指受施者的糖化後期產物的含量降低、或該產物所包含的官能基團的含量改善。Preferably, the inhibiting or slowing down of the recipient's glycation reaction refers to the reduction of the content of the post-glycation product in the recipient, or the improvement of the content of the functional group contained in the product.
較佳地，該糖化後期產物包含高度糖化終產物 （AGEs）。Preferably, the post-glycation products comprise advanced glycation end products (AGEs).
較佳地，該官能基團的含量改善包含羰基的含量降低、ε-胺基的含量提升、游離硫氫基的含量提升。Preferably, the improvement of the content of the functional group includes the reduction of the content of carbonyl groups, the increase of the content of ε-amine groups, and the increase of the content of free sulfhydryl groups.
較佳地，該抑制或減緩受施者的糖化反應係指受施者的高度糖化終產物 （AGEs）與高度糖化終產物受器（RAGE）結合率降低。Preferably, the inhibiting or slowing down of the recipient's glycation response means that the subject's binding rate of advanced glycation end products (AGEs) to receptors for advanced glycation end products (RAGE) is reduced.
較佳地，該皮膚老化的症狀包含皺紋、細紋、斑、鬆弛或其組合。Preferably, the symptoms of skin aging include wrinkles, fine lines, spots, sagging or a combination thereof.
較佳地，該改善皮膚老化包含使受施者的皮膚彈性增加、皮膚水分含量增加或其組合。Preferably, the improving skin aging comprises increasing the recipient's skin elasticity, increasing skin moisture content or a combination thereof.
菌種培養Strain culture
勾取上述短乳酸桿菌的菌落（colony）接種於固態培養基上活化菌種。在一較佳的實施態樣中，該固態培養基為MRS 瓊脂。待菌體生長完成後，將新鮮的菌體連同固態培養基接入於含有液態培養基的錐形瓶中進行液態培養。在一較佳的實施態樣中，於溫度25至40 ℃下、通氣量0至1 vvm氮氣或二氧化碳、速率250至1000 rpm的條件下液態培養。在一較佳的實施態樣中，液態培養的時間為16至24小時，更佳為18小時活菌數可達到最高點。在一較佳的實施態樣中，液態培養基為MRS液態培養基。在一較佳的實施態樣中，液態培養基的配方如下表1所示。液態培養完成後進行發酵培養。在一實施態樣中，發酵培養為有氧或無氧環境。在一實施態樣中，發酵培養的培養溫度控制在30℃至37℃。在一實施態樣中，發酵培養的時間長達16小時至48小時。The above-mentioned Lactobacillus brevis colony (colony) was picked and inoculated on the solid medium to activate the strain. In a preferred embodiment, the solid medium is MRS agar. After the growth of the bacteria is completed, the fresh bacteria and the solid medium are inserted into the Erlenmeyer flask containing the liquid medium for liquid culture. In a preferred embodiment, the culture is carried out in a liquid state at a temperature of 25 to 40° C., an aeration rate of 0 to 1 vvm nitrogen or carbon dioxide, and a rate of 250 to 1000 rpm. In a preferred embodiment, the time for liquid culture is 16 to 24 hours, more preferably 18 hours for the number of viable bacteria to reach the highest point. In a preferred embodiment, the liquid medium is MRS liquid medium. In a preferred embodiment, the formulation of the liquid medium is shown in Table 1 below. After the liquid culture is completed, the fermentation culture is carried out. In one embodiment, the fermentation culture is an aerobic or anaerobic environment. In one embodiment, the culture temperature of the fermentation culture is controlled at 30°C to 37°C. In one embodiment, the time of fermentation culture is as long as 16 hours to 48 hours.
表1
乳酸桿菌發酵物的製備Preparation of Lactobacillus Ferment
將乳酸桿菌經發酵培養而完成生長後，收集包含菌體與培養液的發酵全液進行離心以分離菌泥與上清液。在一較佳的實施態樣中，發酵全液投入15重量%脫脂乳粉靜置1至4小時使菌體凝集成團狀增加回收率。在一較佳的實施態樣中，離心速率為1000至15000 rpm。後述實驗以發酵液離心後取得的上清液作為乳酸桿菌發酵物。本發明的乳酸桿菌的態樣，包括但不限於前述含有菌體與培養液的發酵全液、發酵全液離心而去除上清液後的菌泥、及發酵全液離心而去除菌泥後的上清液。 關於本發明的乳酸桿菌的含量，相對於本發明之含有乳酸桿菌組成物的總量而言，通常可以為0.1重量%以上、較佳為1~50重量%、更佳為5~30重量%、再更佳為10~15重量%之範圍。After the lactobacilli are fermented and grown to complete the growth, the whole fermentation liquid including the bacteria and the culture liquid is collected and centrifuged to separate the bacteria sludge and the supernatant. In a preferred embodiment, the whole fermentation solution is put into 15% by weight skim milk powder and left to stand for 1 to 4 hours to agglomerate the bacterial cells into agglomerates to increase the recovery rate. In a preferred embodiment, the centrifugal speed is 1000 to 15000 rpm. In the following experiments, the supernatant obtained after the fermentation broth was centrifuged was used as the lactobacillus fermentation product. The aspect of the lactobacillus of the present invention includes, but is not limited to, the above-mentioned fermentation whole liquid containing bacteria and culture liquid, the bacteria sludge after the fermentation whole liquid is centrifuged to remove the supernatant, and the fermentation whole liquid centrifugation to remove the bacteria sludge supernatant. The content of the lactic acid bacteria of the present invention can be usually 0.1% by weight or more, preferably 1 to 50% by weight, more preferably 5 to 30% by weight, relative to the total amount of the composition containing lactic acid bacteria of the present invention. , More preferably in the range of 10 to 15% by weight.
具有肌膚轉換改善作用之植物萃取物或化合物Plant extracts or compounds with skin transformation and improvement effects
本發明之組成物中，因應需要而添加後述具有肌膚轉換改善作用之植物萃取物或化合物。藉由添加具有肌膚轉換改善作用之植物萃取物或化合物，使以乳酸桿菌所達到的抗皺紋作用、抗細紋作用、美白作用，甚至是抗斑作用、抗鬆弛作用，都能提高至遠遠超出僅僅只有兩者的相加效果。In the composition of the present invention, the following plant extracts or compounds having skin transformation improvement effects are added as needed. By adding plant extracts or compounds that have the effect of improving skin transformation, the anti-wrinkle effect, anti-fine line effect, whitening effect, and even anti-spot effect and anti-relaxation effect achieved by Lactobacillus can be greatly improved Beyond just the additive effect of the two.
前述具有肌膚轉換改善作用之植物萃取物或化合物方面，舉例如芝麻、地黃、桃、甘草、仙人掌、小麥胚芽、野草莓、鼠尾草、薄荷、胡椒薄荷、魚腥草、鉤藤、紫蘇、琵琶、日本山茶、棗、蓼藍、木通、明日葉、羅漢柏、西印度櫻桃、歐洲蜀葵、蘆薈、銀杏、黃檗、小連翹、橄欖、橙、洋甘菊、榅桲、葡萄柚、桂皮、月桃、地榆、白樺、楊桃、獐牙菜、桑白皮、茶、丁香、胡蘿蔔、 北美金縷梅、薔薇、喜瑪拉雅覆盆子、假葉樹、擬金雀花、牡丹根皮、歐洲七葉樹、月見草、矢車草、桉樹、虎耳草、柚子、薏苡仁、蘭、檸檬、紫雲英、玫瑰果、迷迭香、櫻、鹿角藻、狼毒、金果欖、蛇床子、波爾多樹、蓖麻、檫樹、當歸、矮瑞香、柯氏乳香樹、甘遂、鳳仙花、連翹、紫苜蓿、沒藥、草烏桕、石松、吐金草、芎藭、無花果、木蘭、白木蘭、白樺、大葉藜、忍冬、梔子、桃仁、棗等植物之萃取物、海藻、綠藻、蜂王漿、陳皮。The aforementioned plant extracts or compounds that have the effect of improving skin transformation, such as sesame, rehmannia glutinosa, peach, licorice, cactus, wheat germ, wild strawberry, sage, mint, peppermint, houttuynia cordata, uncaria, perilla , Pipa, Japanese Camellia, Jujube, Polygonum indigo, Akebia, Ashitaba, Luo Hanbo, Acerola, Hollyhock, Aloe Vera, Ginkgo, Phellodendron, Forsythia, Olive, Orange, Chamomile, Quince, Grapefruit, Cinnamon, Moon peach, Burnet, White birch, Carambola, Swertia, Morus alba, Tea, Clove, Carrot, Witch hazel, Rose, Himalayan raspberry, False leaf tree, Gorse, Peony root bark , horse chestnut, evening primrose, cornflower, eucalyptus, saxifrage, grapefruit, coix seed, orchid, lemon, vetch, rosehip, rosemary, cherry, staghorn, chamaema chamaejasme, golden fruit olive, cnidium, Bordeaux Tree, castor, sassafras, angelica, daphne, frankincense, kansui, impatiens, forsythia, alfalfa, myrrh, tallow, stone pine, golden grass, xiongzi, fig, magnolia, white magnolia, Birch, quinoa, honeysuckle, gardenia, peach kernel, jujube and other plant extracts, seaweed, green algae, royal jelly, tangerine peel.
前述具有肌膚轉換改善作用之化合物方面，舉例如肉毒鹼、菸鹼酸醯胺、吡多辛鹽酸鹽(維生素B6)、玻尿酸、甘醇酸、檸檬酸、乳酸、蘋果酸、酒石酸、2-羥基酪酸、3-羥基酪酸、丙醇二酸、維他命A類、甘草酸衍生物、尿囊素、雙氯氛酸、聯苯乙酸、異丁苯丙酸、凱妥普洛芬、羅索普洛芬及該等之鹽。The aforementioned compounds that have the effect of improving skin transformation include carnitine, niacinamide, pyridoxine hydrochloride (vitamin B6), hyaluronic acid, glycolic acid, citric acid, lactic acid, malic acid, tartaric acid, 2 -Hydroxybutyric acid, 3-hydroxybutyric acid, tartronic acid, vitamin A, glycyrrhizic acid derivatives, allantoin, diclofenac, felbinac, ibuprofen, ketoprofen, rosso Profen and its salts.
前述具有肌膚轉換改善作用之植物萃取物之中，係以柏科的植物(羅漢柏等)、銀杏科的植物(銀杏等)、柑橘科的植物(黃檗、橙、葡萄柚、柚子、檸檬等)、紫蘇科的植物(薄荷、胡椒薄荷、紫蘇、迷迭香等)、魚腥草科的植物(魚腥草等)、及杜仲科的植物(杜仲等)之萃取物較適用。又以羅漢柏萃取物、銀杏萃取物、黃檗萃取物、橙萃取物、紫蘇萃取物、魚腥草萃取物及杜仲茶葉萃取物，因具有後述糖化反應抑制作用之故，在本發明中和乳酸桿菌的組合最為適用。Among the above-mentioned plant extracts that have the effect of improving skin transformation, there are plants of Cupressaceae (Tumba chinensis, etc.), plants of Ginkgoceae (Ginkgo biloba, etc.), plants of Citrus family (corkberry, orange, grapefruit, pomelo, lemon, etc.) ), plants of Perillaceae (mint, peppermint, perilla, rosemary, etc.), plants of Houttuyniaceae (Houttuynia cordata, etc.), and extracts of plants of Eucommiaceae (Eucommia, etc.) are more suitable. In addition, extracts of Luo Hanbo, Ginkgo biloba, Phellodendron barkberry, Orange extract, Perilla extract, Houttuynia cordata extract and Eucommia tea extract can neutralize lactic acid in the present invention due to their inhibitory effect on the saccharification reaction described later. Combinations of bacilli are most suitable.
關於前述具有肌膚轉換改善作用之植物萃取物或化合物各自較佳的含量，相對於本發明之含有乳酸桿菌組成物的總量而言，通常可以為0.001重量%以上、較佳為0.01~5重量%、更佳為0.1~20重量%、再更佳為1~10重量%之範圍。With regard to the preferred contents of the above-mentioned plant extracts or compounds with skin transformation improvement effects, relative to the total amount of the composition containing lactobacilli of the present invention, it can usually be more than 0.001% by weight, preferably 0.01 to 5% by weight %, more preferably in the range of 0.1 to 20% by weight, more preferably in the range of 1 to 10% by weight.
具有糖化反應抑制作用之植物萃取物或化合物Plant extracts or compounds with saccharification reaction inhibitory effect
本發明之組成物中，因應需要而添加後述具有糖化反應抑制作用之植物萃取物或化合物。藉由添加具有肌膚轉換改善作用之植物萃取物或化合物，使以乳酸桿菌所達到的抗皺紋作用、抗細紋作用、美白作用，甚至是抗斑作用、抗鬆弛作用，都能提高至遠遠超出僅僅只有兩者的相加效果。In the composition of the present invention, the following plant extract or compound having a saccharification reaction inhibitory effect is added as needed. By adding plant extracts or compounds that have the effect of improving skin transformation, the anti-wrinkle effect, anti-fine line effect, whitening effect, and even anti-spot effect and anti-relaxation effect achieved by Lactobacillus can be greatly improved Beyond just the additive effect of the two.
前述具有糖化反應抑制作用之植物萃取物方面，舉例如菜薊、小米草、透莖冷水麻、小葉胡頹子、龍芽草、荊瀝果實、木通、嫩葉甘藍椰子果實、睡茄、羅漢柏、兒茶、栓皮櫟、絞股藍、尖齒茴香、北美金縷梅、阿里斯艼、淫羊藿、虎杖、銀杏、日本鹿蹄草、草莓、山丹、海蘿、假枇杷、黃鐘木樹皮、紅葉槭、番山丹、茴香、美國金縷梅、矮冬青、鬱金、梅、狹葉櫟、短角淫羊 藿、營實、紫花金光菊、刺五加、金雀花、龍膽、接骨木、微脈冬青、扇椰子、黃檗、黃連、乳薊、燕麥、圓齒碎米薺、豬毛菜、玫瑰百合、黃鵪菜、鬼百合、牛至、橙、南瓜種子、卡圖巴樹皮、鹿子百合、瓜拿納、洛神葵、細辛、熱河蒲公英、桔梗、酸模、海州常山、菲島桐、浙江百合、胡桃、越橘、鴨舌草、佐賀褐藻、石榴、甘藷、撒爾沙根、野山楂、山椒草、紫蘇、翼豆、蚊子草屬、芍藥、掌葉大黃、白山菊、藜、山白蘭、大車前草、積雪草、蕎麥、麝香百合、碎米薺、葡萄藤(Davilla rugosa)、高貴百合、科迪亞柳葉(Cordia salicifolia)、丁香、狹葉番瀉樹、雷公根、雛菊、蒔蘿、吊鏈鉤、山菊、鐵砲百合、吊鏈鉤根、懸鉤子、桃仁、魚腥草、棘梨、杜仲茶葉、蕃茄、洋委陵菜、皺葉酸模、接骨木、旱金蓮科、野紺菊、薄荷、博多百合、胡枝子、大野芋、西番蓮、百香果、巴西人蔘根、菠蘿蜜、緋寒櫻、向日葵、姫蟒蛇草、姫百合、大花黃鵪菜、檳榔、款冬、鵲豆、總狀花升麻、昭和草、木天蓼、香楠、松、馬刀葉椎、馬黛茶、白花百合、小飛雉之朔果、省沽油、野木瓜、海蘊、柳、萱草、山刺番荔枝、山百合、蕹菜、嫁菜、荔枝種子、岷江百合、琉球矢竹、琉球薔薇草莓、蘋果未熟果、椴木花、赤萁、萵苣、檸檬、檸檬香茅、百里香、檸檬葉馬鞭草、香蜂草、玫瑰果、淡紅洋薔薇、迷迭香、短刺野薔薇、月桂、紅景天、墨西哥蘇木、地榆、紫雲英、柿葉、甘草葉、黑大豆種皮、紫米種子、月桃葉、細葉百 合、西洋柳、杜仲葉、明日葉之葉等植物之萃取物、小海蘿、海蘿、鹿角海蘿等之海藻的萃取物、生咖啡豆、甘藷燒酒粕、傘菌屬菌絲體等之萃取物。The above-mentioned plant extracts that have the effect of inhibiting the saccharification reaction, such as artichoke, eyebright, cold water hemp, lobster, asparagus, vitex fruit, akebia, cabbage coconut fruit, nightshade, arhat Cypress, catechin, cork oak, gynostemma, fennel, witch hazel, aristotle, epimedium, knotweed, ginkgo, Japanese wintergreen, strawberry, shandan, seaweed, false loquat, yellow bellwood bark, Red leaf maple, Pansandan, fennel, American witch hazel, dwarf holly, turmeric, plum, oak angustifolia, epimedium shorthorn, camphor, coneflower, eleuthero, gorse, gentian, elderberry Wood, holly, fan palm, amurberry, berberine, milk thistle, oats, catkins, saffron, rose lily, yellow quail, ghost lily, oregano, orange, pumpkin seeds, catuaba bark, Deer lily, guarana, roselle, asarum, Jehol dandelion, platycodon, sorrel, Haizhou Changshan, Philippine island tree, Zhejiang lily, walnut, lingonberry, duck tongue, Saga brown algae, pomegranate, sweet potato, Saar Sand Root, Wild Hawthorn, Sanshope, Perilla, Wing Bean, Mosquito, Peony, Rhubarb palmatum, White Mountain Chrysanthemum, Quinoa, Mountain Pineland, Great Plantain, Centella Asiatica, Buckwheat, Musk Lily, Orina, Grapevine (Davilla rugosa), noble lily, Cordia salicifolia, clove, senna angustifolia, gotu kola, daisy, dill, chain hook, mountain chrysanthemum, iron cannon lily, chain hook root, hanging Hook, peach kernel, houttuynia cordata, prickly pear, Eucommia tea leaves, tomato, Potentilla, sorrel leaf, elderberry, nasturtium, chrysanthemum, mint, Hakata lily, sage, yam, passionflower , passion fruit, Brazilian ginseng root, jackfruit, scarlet cherry, sunflower, Hime boa grass, Hime lily, yellow quail, betel nut, coltsfoot, magpie bean, cohosh raceme, Showa grass, Polygonum chinensis, incense Nan, pine, saber leaf vertebra, yerba mate, white lily, pheasant pheasant fruit, province oil, wild papaya, mozuku, willow, daylilies, soursop, mountain lily, water spinach, weeding vegetables, lychee seeds , Minjiang Lily, Ryukyu Yazhu, Ryukyu Rose Strawberry, Unripe Apple Fruit, Basswood Flower, Osmanthus, Lettuce, Lemon, Lemongrass, Thyme, Lemon Leaf Verbena, Lemon Balm, Rosehip, Pale Red Cinnamon, Myth Rosemary, briar rose, laurel, rhodiola rosea, Mexican sumac, Burnet, milk vetch, persimmon leaf, licorice leaf, black soybean seed coat, purple rice seed, moon peach leaf, lily lily, western willow, Eucommia leaf, tomorrow Extracts of plants such as leaves of leaves, extracts of seaweed such as seaweed, seaweed, and antler seaweed, extracts of green coffee beans, sweet potato shochu, and agaric mycelium.
前述具有糖化反應抑制作用之化合物方面，舉例如雌馬酚、異黃酮、1,4-蒽醌、1-胺基-2-羥基甲基蒽醌、4-胺基苯酚、1,3,5-三羥基苯、麴酸、3,4-二羥基苯基乙酸、咖啡酸、艾芬地爾(ifenprodil)、6-羥基-2,5,7,8-四甲基二氫苯并哌喃-2-羧酸、6-羥基吲哚、7-羥基-4,6-二甲基酞內酯、α-硫辛酸、4-羥基查耳酮、真珠蛋白質水解物、胺基胍、赤藻辛鈉、麥角硫醇、白藜蘆醇、3,3',5,5'-四羥基二苯乙烯等之羥基二苯乙烯類、合氧吲哚、肌肽、水楊酸、去甲豬毛菜鹼溴化氫酸鹽、芥籽酸、菸鹼酸生育酚酯、菸鹼酸醯胺、降二氫癒創木酸、花青素、甘露醇、水解酪蛋白、水解性單寧、兒茶酚、綠原酸、異綠原酸、新綠原酸、隱綠原酸、咖啡醯奎寧酸等之綠原酸類、白矢車菊素、野樺素、前花青二醇寡聚物、葡糖基芸香苷等。The above-mentioned compounds with saccharification reaction inhibitory effect, such as equol, isoflavones, 1,4-anthraquinone, 1-amino-2-hydroxymethylanthraquinone, 4-aminophenol, 1,3,5 -Trihydroxybenzene, kojic acid, 3,4-dihydroxyphenylacetic acid, caffeic acid, ifenprodil, 6-hydroxy-2,5,7,8-tetramethyldihydrobenzopyran -2-Carboxylic acid, 6-oxindole, 7-hydroxy-4,6-dimethylphthalide, α-lipoic acid, 4-hydroxychalcone, pearl protein hydrolyzate, aminoguanidine, red algae Octyl sodium, ergothiol, resveratrol, hydroxystilbenes such as 3,3',5,5'-tetrahydroxystilbene, oxindole, carnosine, salicylic acid, norporcine Ericaine bibromide, sinapic acid, nicotinic acid tocopheryl ester, nicotinic acid amide, nordihydroguaiaretic acid, anthocyanins, mannitol, hydrolyzed casein, hydrolyzed tannin, Catechol, chlorogenic acid, isochlorogenic acid, neochlorogenic acid, cryptochlorogenic acid, caffeoylquinic acid and other chlorogenic acids, cyanidin, betulin, procyanine glycol oligomers, Glucosylrutin, etc.
前述成分之中，睡茄、羅漢柏、兒茶、銀杏、矮冬青、黃檗、橙、南瓜種子、卡圖巴樹皮、菲島桐、葡糖基芸香苷、越橘、紫蘇、蕎麥葉、葡萄藤(Davilla rugosa)、魚腥草、姫蟒蛇草、昭和草、木天蓼、琉球矢竹、琉球薔薇草莓、蘋果未熟果、柿葉、甘草葉、月桃葉、杜仲葉的萃取物、及雌馬酚、異黃酮、艾芬地爾(ifenprodil)、真珠蛋白質水解物、兒茶酚、咖啡酸、及野樺素，因可有效地提高以包含乳酸桿菌的組成物所致抗皺紋作用、抗細紋作用、美白作用，甚至是抗斑作用、抗鬆弛作用，而可作為本發明中更佳的組合。Among the aforementioned ingredients, withania, luohanbo, catechin, ginkgo, dwarf holly, amurberry, orange, pumpkin seeds, catuaba bark, phenanthrene, glucosyl rutin, lingonberry, perilla, buckwheat leaves, grape Rattan (Davilla rugosa), Houttuynia cordata, Hime boa constrictor grass, Showa grass, Polygonum japonica, Ryukyu arrow bamboo, Ryukyu rose strawberry, unripe apple fruit, persimmon leaf, licorice leaf, moon peach leaf, Eucommia leaf extract, and female Equor, isoflavones, ifenprodil, pearl protein hydrolyzate, catechol, caffeic acid, and betulin, because they can effectively improve the anti-wrinkle effect and anti-aging effect caused by the composition containing lactobacillus. Fine-grain effect, whitening effect, even anti-spot effect and anti-relaxation effect can be used as a better combination in the present invention.
前述具有糖化反應抑制作用之植物萃取物之中，又以柏科的植物(羅漢柏等)、銀杏科的植物(銀杏等)、柑橘科的植物(黃檗、橙、檸檬等)、紫蘇科的植物(紫蘇、迷迭香等)、魚腥草科的植物(魚腥草等)及杜仲科的植物(杜仲等)之萃取物較適用。其中，羅漢柏萃取物、銀杏萃取物、黃檗萃取物、橙萃取物、紫蘇萃取物、魚腥草萃取物、及杜仲葉萃取物，因具有肌膚的轉換改善作用，在本發明中，係以與乳酸桿菌的組合最為適用。Among the above-mentioned plant extracts with saccharification reaction inhibitory effect, there are also plants of Cupressaceae (Podocarpus, etc.), plants of Ginkgoceae (Ginkgo, etc.), plants of Citrus (corkberry, orange, lemon, etc.), plants of Perillaceae Extracts of plants (perilla, rosemary, etc.), plants of the Houttuyniaceae family (Houttuynia cordata, etc.), and plants of the Eucommiaceae family (Eucommia, etc.) are more suitable. Among them, Luo Hanbo extract, Ginkgo biloba extract, Phellodendron bark extract, Orange extract, Perilla extract, Houttuynia cordata extract, and Eucommia leaf extract have skin conversion and improvement effects. In the present invention, they are based on Combination with Lactobacillus works best.
將此等具有糖化反應抑制作用之植物萃取物或化合物與乳酸桿菌併用時，因可有效地抑制糖化最終產物(advanced glycation end products：AGEs)的生成，除了可減低乃至於抑制不佳的AGEs對皮膚組織之沈澱附著之外，同時在生成AGEs的過程中，因可在膠原蛋白的分子內及分子間形成無秩序的交聯，並可減低乃至於抑制膠原蛋白中所引起的物理性、生理性變化(改性)，因而得以有效地預防乃至於改善皺紋、肌膚粗糙、暗沈、或緊緻的降低等肌膚的老化。再者，將具有糖化反應抑制作用之植物萃取物與乳酸桿菌併用時，可發揮下列的優點：封鎖起因於該等植物萃取物之異味、異臭，同時更加地提昇保存安定性。When these plant extracts or compounds with saccharification reaction inhibitory effect are used together with Lactobacillus, it can effectively inhibit the formation of saccharification end products (advanced glycation end products: AGEs), in addition to reducing or even poorly inhibiting AGEs on In addition to the precipitation and adhesion of skin tissue, at the same time, in the process of generating AGEs, it can form disordered cross-links in the molecules and between molecules of collagen, and can reduce or even inhibit the physical and physiological effects caused by collagen. Change (modification), so it can effectively prevent or even improve skin aging such as wrinkles, skin roughness, dullness, or reduction of firmness. Furthermore, when the plant extracts having the saccharification reaction inhibitory effect are used together with lactobacilli, the following advantages can be exerted: the odor and odor caused by the plant extracts are blocked, and the storage stability is further improved.
關於前述具有糖化反應抑制作用之植物萃取物或化合物各自較佳的含量，相對於本發明之含有乳酸桿菌組成物的總量而言，通常可以為0.001重量%以上、較佳為0.01~5重量%、更佳為0.1~20重量%、再更佳為1~10重量%之範圍。Regarding the preferred contents of the above-mentioned plant extracts or compounds having a saccharification reaction inhibitory effect, relative to the total amount of the composition containing lactobacilli of the present invention, it can usually be more than 0.001% by weight, preferably 0.01 to 5% by weight. %, more preferably in the range of 0.1 to 20% by weight, more preferably in the range of 1 to 10% by weight.
組成物的具體產品及其添加物Concrete products of the composition and their additives
本發明組成物可透過外用適用於化妝品、彩妝化妝品等之化妝品的領域，特別是添加至直接適用於皮膚的化妝水、洗面劑、美容液、乳液、乳霜等之化妝品時，具有優異的抗老化效果。The composition of the present invention can be applied to the field of cosmetics such as cosmetics and make-up cosmetics through external use, especially when added to cosmetics such as lotion, facial cleanser, beauty lotion, emulsion, and cream that are directly applied to the skin, it has excellent anti-aging properties. aging effect.
化妝水lotion
本發明之組成物，可以水性介質作為基劑，並於其中進一步摻合乙醇及保濕劑而成為化妝水。化妝水係可用以補給水分或保濕成分至皮膚的角質層，同時可用於協助皮膚的生理作用。化妝水舉例如清淨用化妝水、收斂性化妝水、柔軟性化妝水、多層狀化妝水等，也包含肌膚乳液、身體乳液、護手乳液、保濕乳液等。The composition of the present invention can use an aqueous medium as a base, and further blend ethanol and a moisturizing agent into it to form a lotion. The lotion system can be used to supply moisture or moisturizing ingredients to the stratum corneum of the skin, and can also be used to assist the physiological functions of the skin. Lotions include cleansing lotion, astringent lotion, softening lotion, multi-layered lotion, etc., and also include skin lotion, body lotion, hand lotion, moisturizing lotion, and the like.
摻合本發明之組成物所製備的化妝水，係使用保濕效果優異，並組合乳酸桿菌後能更有效地發揮肌膚粗糙改善效果之保濕劑為佳。保濕劑舉例如甘油、乙二醇、丙二醇、1,3-丁二醇、3-甲基-1,3-丁烷二醇、季戊四醇、二甘油、聚甘油、二乙二醇、三乙二醇、聚乙二醇、二丙二醇、海藻糖、神經醯胺類、2-甲基丙烯醯基氧乙基磷膽鹼、玻尿酸、硫酸軟骨素、水解玻尿酸、酵母萃取液、薏苡仁精華液、牡丹精華液、海藻精華液、龍膽精華液、三甲基甘胺酸、N-甲基-L-絲胺酸、菸鹼酸醯胺、蜂王漿精華液。保濕劑相對於本發明之化妝水之總量而言，通常可以為0.0001重量%以上、較佳為0.001~50重量%、更佳為0.01~25重量%、再更佳為0.1~10重量%之範圍下摻合。For the lotion prepared by blending the composition of the present invention, it is better to use a moisturizer that has an excellent moisturizing effect and can more effectively exert the effect of improving rough skin by combining Lactobacillus. Humectants such as glycerin, ethylene glycol, propylene glycol, 1,3-butanediol, 3-methyl-1,3-butanediol, pentaerythritol, diglycerin, polyglycerin, diethylene glycol, triethylene glycol Alcohol, polyethylene glycol, dipropylene glycol, trehalose, ceramides, 2-methacryloxyethylphosphocholine, hyaluronic acid, chondroitin sulfate, hydrolyzed hyaluronic acid, yeast extract, coix seed essence, Peony essence, seaweed essence, gentian essence, trimethylglycine, N-methyl-L-serine, niacinamide, royal jelly essence. The amount of the moisturizer is generally 0.0001% by weight or more, preferably 0.001 to 50% by weight, more preferably 0.01 to 25% by weight, and even more preferably 0.1 to 10% by weight, based on the total amount of the lotion of the present invention blended within the range.
摻合本發明之組成物所製備的化妝水，使用收斂效果優異之收斂劑為佳。收斂劑舉例如檸檬酸、琥珀酸、酒石酸、視黃酸、蘋果酸等之酸劑、視黃醇、氯化鋅、鋁末、硫酸鋁等之鋁化合物、明礬及改性醇、乙醇、歐洲蜀葵精華液、兔菊屬精華液、拳蓼萃取液、咬人貓精華液、鳶尾根精華液、茴香精華液、烏龍茶精華液、乙醇、大花紫薇精華液、小連翹精華液、橙花水、橙果汁、茵陳蒿精華液、木莓精華液、番石榴精華液、黑醋栗果實精華液、香葉醇、牻牛兒苗精華液、野山楂精華液、蚊子草屬精華液、芍藥精華液、白樺精華液、忍冬精華液、問荊精華液、常春藤精華液、西洋野山楂精華液、百里香麝香草精華液、茶精華液、魚腥草精華液、香芹精華液、北美金縷梅精華液、葡萄葉精華液、松精華液、香蜂草精華液、矢車菊精華液、艾屬精華液、檸檬果汁、紫雲英精華液、地榆精華液。收斂劑相對於本發明之化妝水之總重而言，通常為0.0001重量%以上、較佳為0.001~50重量%、更佳為0.01~25重量%、再更佳為0.1~10重量%之範圍下摻合。For the lotion prepared by blending the composition of the present invention, it is better to use an astringent with excellent astringent effect. Astringents include acid agents such as citric acid, succinic acid, tartaric acid, retinoic acid, malic acid, etc., retinol, zinc chloride, aluminum powder, aluminum compounds such as aluminum sulfate, alum and modified alcohols, ethanol, European Hollyhock Essence, Rabbit Chrysanthemum Essence, Polygonum Essence, Biting Cat Essence, Orris Root Essence, Fennel Essence, Oolong Tea Essence, Ethanol, Crape Myrtle Essence, Forsythia Essence, Orange Blossom Water . Essence, white birch essence, honeysuckle essence, quince essence, ivy essence, western wild hawthorn essence, thyme thyme essence, tea essence, houttuynia cordata essence, parsley essence, North American gold Hazel essence, grape leaf essence, pine essence, lemon balm essence, cornflower essence, mugwort essence, lemon juice, vetch essence, burnet elm essence. The astringent is usually 0.0001% by weight or more, preferably 0.001-50% by weight, more preferably 0.01-25% by weight, and even more preferably 0.1-10% by weight, relative to the total weight of the lotion of the present invention. Blended under range.
摻合本發明之組成物所製備的化妝水的，使用角質軟化效果優異之角質軟化劑為佳。角質軟化劑舉例如尿素、硫、乙醇、乳酸、甘醇酸、水楊酸、硫酸鈉等。For the lotion prepared by blending the composition of the present invention, it is preferable to use a cuticle softening agent with excellent cuticle softening effect. Cuticle softening agents include, for example, urea, sulfur, ethanol, lactic acid, glycolic acid, salicylic acid, sodium sulfate, and the like.
軟化(柔軟)劑方面，主要以近似皮脂之油性成分為佳。摻合本發明之組成物所製備的化妝水，使用軟化效果優異，且可泛用於化妝品領域之軟化(柔軟)劑為佳。軟化(柔軟)劑舉例如鯨蠟醇、月桂基醇、鯨蠟硬脂醇、硬脂醯醇、油醇、山崳醇、羊毛脂醇、氫化羊毛脂醇、己基癸醇及辛基十二醇、鯊烷、植物性鯊烷、2-乙基己烷酸十六烷基、α-烯烴寡聚物、異壬酸異十三基、異壬酸異壬基酯、異壬酸鯨蠟、油酸乙基酯、油酸油基、棕櫚酸乙基己基酯、辛醯二醇、氫化大豆磷脂質。In terms of softening (softening) agents, oily components similar to sebum are preferred. For the lotion prepared by blending the composition of the present invention, it is better to use a softener (softening) agent that has excellent softening effect and can be widely used in the field of cosmetics. Softening (softening) agents such as cetyl alcohol, lauryl alcohol, cetearyl alcohol, stearyl alcohol, oleyl alcohol, behenyl alcohol, lanolin alcohol, hydrogenated lanolin alcohol, hexyldecanol and octyldodecyl alcohol Alcohol, squalane, vegetable squalane, cetyl 2-ethylhexanoate, α-olefin oligomer, isotridecyl isononanoate, isononyl isononanoate, cetyl isononanoate , Ethyl Oleate, Oleyl Oleate, Ethylhexyl Palmitate, Capryl Glycol, Hydrogenated Soy Phospholipids.
本發明之組成物所製備的化妝水，使用抑制化妝品的經時性混濁、沈澱之效果高，且泛用於化妝品領域之界面活性劑者為佳。界面活性劑舉例如L-精胺酸乙基．DL-吡咯啶酮羧酸鹽(L-arginine ethyl⋅DL-pyrrolidone carboxylic acid salt)、焦麩胺酸異硬脂酸聚氧乙烯甘油酯、PEG-9聚二甲基矽氧基乙基二甲基矽氧烷、椰子油脂肪酸山梨醇酐、聚氧乙烯聚氧丙烯癸基十四基醚、聚氧乙烯十六烷基醚、聚氧乙烯硬脂醯醚、聚氧乙烯甲基葡萄糖苷、聚氧乙烯月桂基醚、聚氧乙烯硬化蓖麻籽油、聚氧乙烯氫化蓖麻籽油、異硬脂酸山梨醇酐、異硬脂酸聚氧乙烯硬化蓖麻籽油、甘油脂肪酸酯、二異硬脂酸聚甘油基、蔗糖脂肪酸酯、硬脂醯基麩胺酸鈉、硬脂醯基乳酸鈉、倍半異硬脂酸山梨醇酐、倍半油酸山梨醇酐、聚氧乙烯烷基(C12-15)醚磷酸、聚氧乙烯膽固醇基 醚、聚氧乙烯十六烷基醚磷酸、聚氧乙烯十六硬脂醯醚、聚氧乙烯植物固醇、聚氧乙烯山崳醇醚、單油酸聚氧乙烯山梨醇酐(20E.O)、單硬脂酸甘油、單硬脂酸聚乙二醇酯、單硬脂酸聚氧乙烯甘油酯、單月桂酸聚氧乙烯山梨醇酐、月桂醯基麩胺酸二辛基十二基酯、水解膠原蛋白液、單硬脂酸甘油酯、氫化大豆溶血磷脂質、氫化大豆磷脂質、及水溶性膠原蛋白。For the lotion prepared from the composition of the present invention, it is preferable to use a surfactant that has a high effect of inhibiting the turbidity and precipitation of cosmetics over time and is widely used in the field of cosmetics. Surfactants such as L-arginine ethyl. DL-pyrrolidone carboxylate (L-arginine ethyl⋅DL-pyrrolidone carboxylic acid salt), polyoxyethylene glyceryl pyrroglutamic acid isostearate, PEG-9 polydimethylsiloxy ethyl dimethyl Silicone, coconut oil fatty acid sorbitan, polyoxyethylene polyoxypropylene decyl tetradecyl ether, polyoxyethylene cetyl ether, polyoxyethylene stearyl ether, polyoxyethylene methyl glucoside, Polyoxyethylene lauryl ether, polyoxyethylene hardened castor oil, polyoxyethylene hydrogenated castor oil, sorbitan isostearate, polyoxyethylene isostearate hardened castor oil, fatty acid esters of glycerol , polyglyceryl diisostearate, sucrose fatty acid ester, sodium stearyl glutamate, sodium stearyl lactate, sorbitan sesquiisostearate, sorbitan sesquioleate, polyoxygen Vinyl alkyl (C12-15) ether phosphoric acid, polyoxyethylene cholesteryl ether, polyoxyethylene cetyl ether phosphoric acid, polyoxyethylene cetearyl ether, polyoxyethylene phytosterol, polyoxyethylene mountain Alcohol ether, polyoxyethylene sorbitan monooleate (20E.O), glycerin monostearate, polyethylene glycol monostearate, polyoxyethylene glycerol monostearate, polyoxyethylene monolaurate Sorbitan, dioctyldodecyl laurylglutamate, hydrolyzed collagen solution, glyceryl monostearate, hydrogenated soybean lysophospholipids, hydrogenated soybean phospholipids, and water-soluble collagen.
再者，為了提昇化妝水的品質與安定性，可摻合香料、色素、防腐劑、殺菌劑、紫外線吸收劑、金屬封鎖劑及緩衝劑。該等物質相對於本發明之化妝水總量而言，通常為0.0001重量%以上、較佳為0.001~50重量%、更佳為0.01~25重量%、再更佳0.1~10重量%之範圍下摻合。Furthermore, in order to improve the quality and stability of the lotion, fragrances, pigments, preservatives, bactericides, ultraviolet absorbers, metal blocking agents and buffers can be blended. These substances are usually 0.0001% by weight or more, preferably 0.001 to 50% by weight, more preferably 0.01 to 25% by weight, and more preferably 0.1 to 10% by weight, relative to the total amount of the lotion of the present invention. Blend down.
洗面劑facial cleanser
本發明之皮膚外用組成物乃藉由摻合具有洗淨作用之成分，而得以作為洗面劑。對皮膚上的髒污而言，係有皮膚本來的生理作用所產生的垢(dirty)，意即，皮脂膜酸敗、變質者與角質層已剝離者之外，尚有空氣中的灰塵等之髒污混然成一體者及日常使用的化妝品，洗面劑乃是為了將此等洗掉而保持皮膚清淨之故幾乎可每日使用。洗面劑的具體例方面，可舉出皂、身體洗劑、洗面乳霜、彩妝去除劑等。具有洗淨作用之成分方面，可舉出高級脂肪酸的鹼鹽(皂)，且高級脂肪酸的鹼鹽主要為C10~C18脂肪酸的鈉鹽、鉀鹽、胺鹽。The composition for external use on skin of the present invention can be used as a facial cleanser by blending ingredients having a cleansing effect. As far as the dirt on the skin is concerned, it is the dirt (dirty) produced by the original physiological function of the skin, that is, in addition to the rancidity and deterioration of the sebum membrane and those whose cuticle has been peeled off, there are also dust in the air, etc. Dirt and daily use of cosmetics, facial cleanser can be used almost every day in order to wash away these dirt and keep the skin clean. Specific examples of face cleansers include soaps, body washes, face cleansers, and make-up removers. In terms of ingredients having a cleansing effect, alkali salts (soaps) of higher fatty acids can be mentioned, and the alkali salts of higher fatty acids are mainly sodium salts, potassium salts, and amine salts of C10~C18 fatty acids.
洗面劑中，為了品質的提升與安定化，可進一步摻合由香料、色素、抗氧化劑及金屬封鎖劑選出的1種或2種以上，該等物質相對於本發明之洗面劑總重量而言，通常為0.0001重量%以上、較佳為0.001~50重量%、更佳為0.01~25重量%、再更佳0.1~10重量%之範圍下摻合。洗面劑中，可進一步摻合由甘油、乙醇、砂糖、多元醇及殺菌劑選出的1種或2種以上。殺菌劑在洗面劑等皂系之洗淨劑要達到消毒、殺菌、防臭之目的下，係以使用鹽酸克羅西希定、異丙基甲基苯酚、感光素101號、甲苯酚、氯二甲酚、三氯羰基胺苯、鹵卡班、苯酚、氯甲苯酚、苯氧基乙醇、三氯羥基二苯基醚等為佳。In the facial cleanser, in order to improve and stabilize the quality, one or more selected from fragrances, pigments, antioxidants and metal blocking agents can be further blended. These substances are relative to the total weight of the facial cleanser of the present invention. , usually at least 0.0001% by weight, preferably 0.001 to 50% by weight, more preferably 0.01 to 25% by weight, and more preferably 0.1 to 10% by weight. In the face cleanser, one or two or more selected from glycerin, ethanol, sugar, polyhydric alcohols, and germicides may be further blended. Bactericides In soap-based detergents such as facial cleansers to achieve the purpose of disinfection, sterilization, and deodorization, the use of clothexidine hydrochloride, isopropyl methylphenol, photosensitive element 101, cresol, and chlorine dichloride Cresol, trichlorocarbonylaminobenzene, halocarban, phenol, chlorocresol, phenoxyethanol, trichlorohydroxydiphenyl ether, etc. are preferred.
美容液Beauty Essence
本發明之皮膚外用組成物，乃藉由摻合保濕劑及柔軟劑，可作為美容液。一般而言，所謂美容液係與化妝水不同，是具有黏度且使用感及保濕性優異的型態之化妝品，被認為具有保濕機能與軟化機能者。劑型已知有可溶化型、O/W乳化型、油液型。The composition for external use on skin of the present invention can be used as a beauty essence by blending a moisturizing agent and a softening agent. Generally speaking, the so-called beauty essence is different from lotion, and it is a type of cosmetic that has viscosity, excellent usability and moisturizing properties, and is considered to have moisturizing and softening functions. Known dosage forms include dissolvable type, O/W emulsified type, and oily liquid type.
一般而言，美容液大多是可藉由進一步摻合由香料、紫外線吸收劑、美白劑、抗發炎劑、防腐劑及殺菌劑選出的1種或2種以上，而具有保濕機能與軟化機能以外的機能者。抗發炎劑較佳係以單獨或組合使用來自天然成分的甘草酸硬脂醯、甘草酸二鉀等之甘草酸或其衍生物、龍膽精華液、紫蘇精華液、鼠尾草精華液、西洋接骨木精華液、雷公根精華液、魚腥草精華液、當歸精華液、牡丹精華液、月見草精華液、無患子精華液、虎耳草精華液、艾屬精華液、蘋果精華液之1種或2種以上為佳。該等物質相對於本發明之美容液總量而言，通常為0.0001重量%以上、較佳為0.001~50重量%、更佳為0.01~25重量%、再更佳0.1~10重量%之範圍下摻合。In general, most of the beauty essences have a moisturizing function and a softening function by further blending one or more kinds selected from fragrances, ultraviolet absorbers, whitening agents, anti-inflammatory agents, preservatives, and bactericides. the functional person. The anti-inflammatory agent is preferably used alone or in combination with glycyrrhizic acid or its derivatives such as stearyl glycyrrhizinate and dipotassium glycyrrhizinate, gentian essence, perilla essence, sage essence, Western Elderberry essence, gotu kola essence, Houttuynia cordata essence, angelica essence, peony essence, evening primrose essence, Sapinberry essence, saxifrage essence, mugwort essence, apple essence or one 2 or more are preferred. These substances are usually in the range of 0.0001% by weight or more, preferably 0.001-50% by weight, more preferably 0.01-25% by weight, and more preferably 0.1-10% by weight, relative to the total amount of the beauty essence of the present invention Blend down.
乳液lotion
本發明之皮膚外用組成物，係以水性介質作為基劑者，並藉由摻合油性成分及界面活性劑，而得以成為乳液。乳液之使用目的在於皮膚的清潔、卸妝、皮膚的保護、保濕及柔軟、以及皮膚的血行促進。乳液除了特別的情況外，乃因油相量較乳霜少而為具流動性之乳劑，故別名亦稱為「液體乳霜」。乳化之型有O/W型及W/O型之兩者，但一般而言為O/W型，因水相之比率多，塗佈於皮膚時，可輕薄延伸而賦予清爽之感觸。油性成分及界面活性劑方面，可適當地使用前述化妝水之項中詳述的各種成分。The composition for external use on the skin of the present invention uses an aqueous medium as a base, and can be formed into an emulsion by blending an oily component and a surfactant. The purpose of the lotion is to clean the skin, remove makeup, protect the skin, moisturize and soften it, and promote the blood circulation of the skin. Except for special circumstances, emulsion is a fluid emulsion because the amount of oil phase is less than that of cream, so it is also called "liquid cream". There are two types of emulsification: O/W type and W/O type, but generally speaking, it is O/W type. Because the ratio of water phase is large, when applied to the skin, it can be stretched lightly and give a refreshing feeling. For the oily component and surfactant, various components described in detail in the section of the aforementioned lotion can be used appropriately.
本發明之乳液中，係可進一步因應目的與機能，來摻合由保濕劑、柔軟劑、血行促進劑及高分子物質選出的1種或2種以上，該等通物質相對於本發明之乳液總量而言，通常為重0.0001重量%以上、較佳為0.001~50重量%、更佳為0.01~25重量%、再更佳為0.1~10重量%之範圍下摻合。一般而言，乳液多是以保護膠質之目的於水相中預先添加高分子物質，高分子物質可舉出羧基乙烯基聚合物、三仙膠、丙烯酸聚合物、羧基甲基纖維素鈉(CMC)等。In the emulsion of the present invention, one or two or more selected from moisturizing agents, softeners, blood flow promoters and polymer substances can be blended in accordance with the purpose and function. These substances are compared with the emulsion of the present invention The total amount is usually at least 0.0001% by weight, preferably 0.001 to 50% by weight, more preferably 0.01 to 25% by weight, and more preferably 0.1 to 10% by weight. Generally speaking, emulsions are mostly pre-added polymer substances in the water phase for the purpose of protecting colloids. Examples of polymer substances include carboxyvinyl polymers, sanxian gum, acrylic acid polymers, sodium carboxymethyl cellulose (CMC) )Wait.
乳霜cream
本發明之皮膚外用組成物，係以水性介質作為基劑者，並藉由摻合油性成分及界面活性劑，而得以成為乳霜。乳化的型係與乳液的情況同樣地，有O/W型及W/O型兩者，但乳霜一般因油相量較乳液多，故為無流動性之乳劑。乳霜的使用目的係與乳液同樣地，在於皮膚的清潔、卸妝、皮膚的保護、保濕及柔軟、以及皮膚的血行促進。更具體而言，可舉出清潔乳霜、軟化乳霜、滋養乳霜、晚霜、底霜、消散乳霜、保濕乳霜、按摩乳霜、冷霜、護唇霜等。The composition for external use on skin of the present invention uses an aqueous medium as a base, and can be formed into a cream by blending oily components and surfactants. The type of emulsification is the same as that of the emulsion, there are both O/W type and W/O type, but the cream is generally a non-fluid emulsion because the oil phase is more than the emulsion. The purpose of the cream is the same as that of the emulsion, which is to clean the skin, remove makeup, protect the skin, moisturize and soften it, and promote the blood circulation of the skin. More specifically, cleansing cream, softening cream, nourishing cream, night cream, primer, dissipating cream, moisturizing cream, massage cream, cold cream, lip care cream, etc. are mentioned.
乳霜中使用的油性成分及界面活性劑，可適用同前述化妝水的各種成分。乳霜係可進一步摻合由鯨蠟、鯨蠟醇、羊毛脂、流動石蠟、凡士林、甘油及鯊烷選出的1種或2種以上作為油性成分之補助成分，該等物質相對於本發明之乳霜總量而言，通常為0.0001重量%以上、較佳為0.001~50重量%、更佳為0.01~25重量%、再更佳為0.1~10重量%之範圍下摻合。The oil-based ingredients and surfactants used in the cream can be applied to the same ingredients as the aforementioned lotion. The cream system can further blend one or more kinds selected from cetyl wax, cetyl alcohol, lanolin, liquid paraffin, vaseline, glycerin and squalane as supplementary ingredients of oily ingredients. The total amount of cream is usually 0.0001% by weight or more, preferably 0.001 to 50% by weight, more preferably 0.01 to 25% by weight, and more preferably 0.1 to 10% by weight.
其他成分other ingredients
本發明之組成物可應用於化妝水、洗面劑、美容液、乳液或者是乳霜中，亦可因應需要而於此等中適當地添加一般所用的其他成分，例 如增黏劑、維他命類、胺基酸類、抗皺紋劑、海藻萃取物、細胞賦活劑、經皮吸收促進劑、發泡劑、可溶化劑、角質溶解劑、賀爾蒙類、顏料、可塑劑、無機粉末、有機粉末等。The composition of the present invention can be applied to lotion, facial cleanser, beauty lotion, lotion or cream, and other commonly used ingredients such as thickeners, vitamins, Amino acids, anti-wrinkle agents, seaweed extracts, cell activators, transdermal absorption accelerators, foaming agents, solubilizers, keratolytic agents, hormones, pigments, plasticizers, inorganic powders, organic powders, etc. .
本發明之組成物，亦可透過內用適用於醫藥或飲食等領域，特別是添加至直接適用於口服的藥品、飼料、飲料、營養補充品、乳製品、食品或保健食品等時，具有優異的抗老化效果。The composition of the present invention can also be used in the fields of medicine or diet through internal use, especially when added to medicines, feeds, beverages, nutritional supplements, dairy products, food or health foods that are directly suitable for oral administration, it has excellent anti-aging effect.
本發明之組合物，可根據用途因應所需要的產品型態以進一步包含添加劑。該添加劑可為賦型劑、防腐劑、稀釋劑、填充劑、吸收促進劑、或甜味劑。該賦型劑可選自檸檬酸鈉、碳酸鈣、磷酸鈣、蔗糖或其組合。該防腐劑可延長組合物的儲藏期限，例如苯甲醇、對羥基苯甲酸（parabens）。稀釋劑可選自水、乙醇、丙二醇、甘油或其組合。填充劑可選自乳糖、高分子量聚乙二醇或其組合。吸收促進劑可選自二甲基亞碸（DMSO）、月桂氮卓酮、丙二醇、甘油、聚乙二醇或其組合。甜味劑可選自安塞甜（Acesulfame K）、阿斯巴甜（aspartame）、糖精（saccharin）、三氯蔗糖／蔗糖素（sucralose）、紐甜（neotame）或其組合。除上述所列舉的添加劑以外，在不影響組合物的醫藥效果前提下，可依需求選用適合的其他添加劑。The composition of the present invention may further contain additives according to the desired product form according to the application. The additive may be an excipient, preservative, diluent, filler, absorption enhancer, or sweetener. The excipient may be selected from sodium citrate, calcium carbonate, calcium phosphate, sucrose or combinations thereof. The preservatives prolong the shelf life of the composition, eg benzyl alcohol, parabens. The diluent may be selected from water, ethanol, propylene glycol, glycerin or combinations thereof. Fillers may be selected from lactose, high molecular weight polyethylene glycols or combinations thereof. The absorption enhancer may be selected from dimethylsulfoxide (DMSO), laurocapram, propylene glycol, glycerin, polyethylene glycol, or combinations thereof. The sweetener may be selected from Acesulfame K, aspartame, saccharin, sucralose, neotame or combinations thereof. In addition to the above-listed additives, other suitable additives can be selected as required, provided that the medicinal effect of the composition is not affected.
藥物型態在不影響組成物所欲發揮的效果之前提，可根據受施予者之需要調製為例如粉劑、錠劑、栓劑、微膠囊、安瓶（ampoule/ampule）、液劑噴劑或塞劑。On the premise that the drug form does not affect the desired effect of the composition, it can be prepared as powder, lozenge, suppository, microcapsule, ampule (ampoule/ampule), liquid spray or Suppository.
施予途徑根據該組合物的型態及受施予者所適合的劑量，舉例如經口或非經腸，還包括靜脈內、肌內、腹膜內、皮下、經皮、呼吸道（氣霧劑）、直腸、***或局部（包括口腔及舌下）。施予的時間點根據該組合物的型態及受施予者所適合的劑量，舉例如每餐、每天、每週。受施予者為人類或動物。The route of administration is based on the form of the composition and the appropriate dose for the recipient, for example, oral or parenteral, and also includes intravenous, intramuscular, intraperitoneal, subcutaneous, transdermal, respiratory (aerosol) ), rectal, vaginal or topical (including oral and sublingual). The timing of administration depends on the form of the composition and the appropriate dose for the recipient, for example, every meal, every day, every week. The recipient is a human or an animal.
糖化反應的實驗設計Experimental design of saccharification reaction
糖化反應的實驗選用葡萄糖 （glucose） 作為糖化分子、甲基乙二醛 （methylglyoxal, MG）、與哺乳動物體內循環系統中含量最高的白蛋白之小牛血清白蛋白 （bovine serum albumin, BSA）分別於以下不同實驗中來進行反應。葡萄糖、甲基乙二醛、小牛血清白蛋白的濃度分別為10 mM、25 mM及5 mg/mL。作為乳酸桿菌所選用的菌株可購自BCRC如下表2，並以前述方法製備乳酸桿菌發酵物。以下的體外實驗結果皆以三重複來計算。Glucose (glucose) was used as the glycation molecule, methylglyoxal (MG), and bovine serum albumin (BSA), the albumin with the highest content in the mammalian circulatory system, were used in the experiment of saccharification reaction. The reactions were carried out in the following different experiments. The concentrations of glucose, methylglyoxal, and bovine serum albumin were 10 mM, 25 mM, and 5 mg/mL, respectively. The strains selected as Lactobacillus can be purchased from BCRC as shown in Table 2, and the Lactobacillus fermented product was prepared by the aforementioned method. The following in vitro experimental results are calculated in triplicate.
表2
實驗一：糖化前期產物的測定Experiment 1: Determination of early glycation products
選用葡萄糖 （glucose） 作為糖化分子與小牛血清白蛋白 （bovine serum albumin, BSA）製備糖化反應液。糖化反應共進行5天，反應溫度控制在37℃。本實驗中的對照組為含糖化分子之BSA，實驗組則是含糖化分子之BSA再加入乳酸桿菌發酵物，以對比糖化BSA（Glycated BSA）或乳酸桿菌發酵物介入的含糖化分子之BSA。取用糖化反應第一天反應液，參考1987年Armbruster等人的方法，利用硝基藍四氮唑(nitroblue tetrazolium，NBT) 試劑測定糖化前期阿馬道里 (Amadori)產物含量。Glucose was selected as the glycosylated molecule and bovine serum albumin (BSA) to prepare the saccharification reaction solution. The saccharification reaction was carried out for 5 days in total, and the reaction temperature was controlled at 37°C. In this experiment, the control group was BSA containing glycosylated molecules, and the experimental group was BSA containing glycosylated molecules added to the fermentation product of Lactobacillus to compare glycosylated BSA (Glycated BSA) or BSA containing glycosylated molecules intervened in the fermentation product of Lactobacillus. The first day of saccharification reaction was taken, and the content of Amadori products in the early stage of saccharification was determined by using nitroblue tetrazolium (NBT) reagent with reference to the method of Armbruster et al. in 1987.
圖2為乳酸桿菌發酵物對蛋白質糖化前期阿馬道里產物生成之影響。對照組為作為糖化蛋白的糖化小牛血清白蛋白(Glycated BSA) 在未添加任何乳酸桿菌發酵物的狀況下，所測得作為阿馬道里產物的酮胺(ketoamine)的濃度為10.72 nmoles/mg，標示如圖中偏右側的垂直線。其他添加乳酸桿菌發酵物的組和對照組相比之下，在糖化前期產生的酮胺的濃度皆降低，大部分介於 2 到 9 nmoles/mg之間。其中編號5和6的乳酸桿菌 GKM3和GKK2的發酵物，分別降低酮胺的濃度至2.82 nmoles/mg和3.47 nmoles/mg。此結果說明本發明的乳酸桿菌或其發酵物，皆有助於抗糖化反應中糖化前期阿馬道里產物生成。Figure 2 shows the effect of Lactobacillus fermentation on the formation of Amadori products in the early stage of protein saccharification. The control group was glycosylated bovine serum albumin (Glycated BSA) as a glycosylated protein. The concentration of ketoamine (ketoamine) as a product of Amadori was measured to be 10.72 nmoles/mg without adding any lactobacillus fermentation product , marking the vertical line to the right of the figure. Compared with the control group, the other groups supplemented with Lactobacillus fermentation products had lower concentrations of ketamines produced in the early stage of glycation, most of which were between 2 and 9 nmoles/mg. The fermentation products of Lactobacillus GKM3 and GKK2 numbered 5 and 6 respectively reduced the concentration of ketamine to 2.82 nmoles/mg and 3.47 nmoles/mg. This result shows that the Lactobacillus of the present invention or its fermented product are all helpful to the formation of amadori products in the early glycation stage in the anti-glycation reaction.
實驗二：糖化中期雙羰基化合物之捕捉效力的評估Experiment 2: Evaluation of the capture efficiency of dicarbonyl compounds in the mid-saccharification stage
混合1mL乳酸桿菌發酵物與1mL濃度為2mM的甲基乙二醛(MG)，並於37℃中反應1小時。再加入1mL濃度12Mm的鄰苯二胺 (o-phenylenediamine, OPD)，於37℃中反應15分鐘，使MG衍生化為2-甲基喹喔啉 (2-methylquinoxaline, 2-MQ)，接著以14000×g離心五分鐘。本實驗中的對照組僅含有MG，實驗組則是MG再加入乳酸桿菌發酵物，以對比MG或加入乳酸桿菌發酵物的MG。參考2011年Lo等人的方法進行HPLC分析，求得乳酸桿菌發酵物捕捉糖化中期雙羰基化合物 MG 之能力。1 mL of the lactobacillus fermentation product was mixed with 1 mL of methylglyoxal (MG) at a concentration of 2 mM, and reacted at 37° C. for 1 hour. Then add 1mL o-phenylenediamine (OPD) with a concentration of 12Mm, and react at 37°C for 15 minutes to derivatize MG into 2-methylquinoxaline (2-MQ), followed by Centrifuge at 14,000 xg for five minutes. In this experiment, the control group contained only MG, and the experimental group was MG added with Lactobacillus ferment to compare MG or MG added with Lactobacillus ferment. Referring to the method of Lo et al. in 2011, HPLC analysis was carried out to determine the ability of Lactobacillus fermentation to capture the dicarbonyl compound MG in the mid-saccharification stage.
圖3為乳酸桿菌發酵物對蛋白質糖化中期雙羰基化合物之捕捉能力評估。將未加入任何乳酸桿菌發酵物之MG 與添加乳酸桿菌發酵物的組相比，以未加入任何乳酸桿菌發酵物之MG顯示數值0作為基準，添加乳酸桿菌發酵物組之捕捉MG能力的數值越高，顯示捕捉MG能力越強。各乳酸桿菌發酵物的組別的數值皆有明顯提升，其中分別編號5和6的乳酸桿菌GKM3、GKK2的發酵物的數值更高而表現出較佳地捕捉MG能力。此結果說明本發明的乳酸桿菌或其發酵物，皆有助於抗糖化反應中糖化中期雙羰基化合物之捕捉。Figure 3 is an evaluation of the ability of Lactobacillus fermentation to capture dicarbonyl compounds in the middle stage of protein saccharification. Comparing the MG without adding any lactobacillus fermentation product with the group adding lactobacillus fermentation product, taking the MG display value of 0 without adding any lactobacillus fermentation product as a benchmark, the value of the MG capture ability of the adding lactobacillus fermentation product group is higher The higher the value, the stronger the ability to capture MG. The values of each group of Lactobacillus fermentation products were significantly improved, and the values of the fermentation products of Lactobacillus GKM3 and GKK2 respectively numbered 5 and 6 were higher, showing better MG capture ability. This result shows that the Lactobacillus or its fermented product of the present invention is helpful for the capture of dicarbonyl compounds in the mid-glycation stage in the anti-glycation reaction.
實驗experiment 三three ：糖化後期糖化程度與其官能基的評估: Evaluation of the degree of saccharification and its functional groups in the late stage of saccharification
本實驗中的對照組為不含糖化分子之BSA、含糖化分子之BSA，實驗組則是含糖化分子之BSA再加入乳酸桿菌發酵物，以對比糖化BSA（Glycated BSA）。取用如實驗一中所配置的糖化反應液，在反應結束後第五天反應液，參考2017年Kumar等人的方法，利用十二烷基硫酸鈉聚丙烯醯胺凝膠電泳 (sodium dodecyl sulfate polyacrylamide gel electrophoresis, SDS-PAGE)分析各乳酸桿菌發酵物組別對蛋白質糖化程度的影響。並分別參考1990年Levine等人的方法，以2,4-二硝基苯肼 (2,4-dinitrophenylhydrazine, DNPH)測定糖化蛋白中的特定官能基羰基含量 (carbonyl content) ；參考2013年Hashimoto等人的方法，以2,4,6-三硝基苯磺酸 (2, 4, 6-trinitrobenzenesulphonic acid, TNBS)測定糖化蛋白中的特定官能基ε-胺基團 (free ε-amino groups)減少含量；以及參考1959年Ellman的方法，以Ellman's reagent (即5,5'-二硫代双(2-硝基苯甲酸)，5,5'-dithiobis-(2-nitrobenzoic acid), DTNB) 進行游離硫氫基團 (free sulfhydryl groups)含量變化測定。In this experiment, the control group was BSA without glycosylated molecules and BSA with glycosylated molecules, and the experimental group was BSA with glycosylated molecules added with Lactobacillus fermentation products to compare glycosylated BSA (Glycated BSA). Take the saccharification reaction solution as configured in
圖4係乳酸桿菌發酵物對蛋白質糖化後期產物之SDS-PAGE圖譜。圖中左欄顯示糖化蛋白 (Glycated BSA) 與未糖化前的蛋白 (BSA)相比，其分子量顯著變大、條帶分布較擴散且集中度低。這是因為蛋白質在糖化過程中，蛋白質的官能基團會產生交聯現象所導致。圖中右側上欄顯示糖化蛋白添加乳酸桿菌發酵物的組別，右側下欄則顯示未糖化前的蛋白添加乳酸桿菌發酵物。右側下欄顯示未糖化前的蛋白添加乳酸桿菌發酵物和左欄未糖化前的蛋白相比，兩者的條帶分布都擴散小、集中度高且差異不明顯。此結果表示這兩組別由於蛋白質都沒有糖化，因此不論是否加入乳酸桿菌發酵物都不會使蛋白質產生交聯現象。圖中右側上欄顯示糖化蛋白添加乳酸桿菌發酵物的組別和左欄糖化蛋白相比之下，SDS-PAGE圖譜中所觀察到添加乳酸桿菌發酵物的糖化蛋白擴散程度都較小且集中程度較高，顯示糖化蛋白的交聯情況均減少。尤其是分別編號5和6的乳酸桿菌GKM3、GKK2的組別，條帶分布擴散小、集中度高，且明顯集中下側，判斷分子交聯 (cross-link)或聚合 (aggregation)的程度更小。此結果說明本發明之菌株皆有助於抑制糖化反應後期蛋白質糖化衍生之交聯聚合反應。Fig. 4 is the SDS-PAGE profile of the late product of protein saccharification by the fermentation product of Lactobacillus. The left column of the figure shows that the glycosylated protein (Glycated BSA) has a significantly larger molecular weight, more diffuse band distribution, and a lower concentration than the unglycosylated protein (BSA). This is because the functional groups of the protein will be cross-linked during the saccharification process of the protein. The upper right column in the figure shows the group in which glycosylated protein is added with lactobacillus fermented product, and the lower column on the right shows the group without saccharified protein added with lactobacillus fermented product. The lower column on the right shows that the protein before unsaccharification is added with Lactobacillus fermentation product and the left column is compared with the protein before unsaccharification. Both band distributions have small diffusion, high concentration and no obvious difference. The results indicated that the proteins in these two groups were not glycosylated, so no matter whether the Lactobacillus fermentation product was added or not, the protein would not be cross-linked. The upper right column of the figure shows the group of glycosylated proteins added with Lactobacillus fermented products, compared with the group of glycosylated proteins added with Lactobacillus fermented products in the SDS-PAGE pattern. Higher, showing that the cross-linking of glycosylated proteins is reduced. Especially for the groups of Lactobacillus GKM3 and GKK2 numbered 5 and 6 respectively, the distribution of the bands has a small diffusion and a high degree of concentration, and is obviously concentrated on the lower side, and the degree of molecular cross-linking or aggregation is more judged. small. This result shows that the strains of the present invention all help to inhibit the cross-linking polymerization reaction derived from protein glycation in the late stage of glycation reaction.
圖5係乳酸桿菌發酵物對蛋白質糖化後期產物羰基含量之影響。在對照組中，BSA (5 mg/mL) 與葡萄糖 (10 mM)、MG (25 mM) 共同反應後，Glycated BSA 的羰基含量增加至 69.74 nmoles/mg ，標示如圖中偏右側的垂直線。反觀加入乳酸桿菌發酵物的組別，羰基增加含量皆低於對照組的含量，而介於25至65nmoles/mg之間。其中編號5和6的乳酸桿菌GKM3、GKK2的發酵物，增加較少的羰基，甚至達30nmoles/mg以下。此結果說明本發明的乳酸桿菌或其發酵物，有助於抗糖化反應後期產物羰基含量的減少。Figure 5 shows the effect of Lactobacillus fermentation product on the carbonyl content of the late product of protein saccharification. In the control group, after co-reaction of BSA (5 mg/mL) with glucose (10 mM) and MG (25 mM), the carbonyl content of Glycated BSA increased to 69.74 nmoles/mg, indicated by the vertical line on the right side of the graph. On the other hand, in the group added with Lactobacillus fermentation product, the increase in carbonyl content was lower than that of the control group, and ranged from 25 to 65 nmoles/mg. Among them, the fermentation products of Lactobacillus GKM3 and GKK2 numbered 5 and 6 increased less carbonyl groups, even reaching below 30nmoles/mg. This result shows that the lactobacillus or its fermented product of the present invention is helpful to resist the reduction of the carbonyl content of the late product of the saccharification reaction.
圖6係乳酸桿菌發酵物對蛋白質糖化後期產物ε-胺基含量之影響。在對照組中，BSA (5 mg/mL) 與葡萄糖(10 mM)、MG (25 mM) 共同反應後，Glycated BSA 的ε-胺基含量減少了 37.28% ，標示如圖中偏中間側的垂直線。反觀加入乳酸桿菌發酵物的部分組別，ε-胺基減少量有改善。其中編號5和6的乳酸桿菌GKM3、GKK2的發酵物，相對於其他種菌株，其降低最多ε-胺基的減少量。此結果說明本發明的乳酸桿菌或其發酵物，有助於改善抗糖化反應後期中的ε-胺基含量減少的現象。Figure 6 shows the effect of lactobacillus fermentation product on the content of ε-amine group in the late product of protein saccharification. In the control group, after the co-reaction of BSA (5 mg/mL) with glucose (10 mM) and MG (25 mM), the ε-amine content of Glycated BSA decreased by 37.28%, indicated by the vertical line on the middle side of the figure Wire. In contrast, the reduction of ε-amine group was improved in some groups added with Lactobacillus fermentation product. Among them, the fermentation products of Lactobacillus GKM3 and GKK2 numbered 5 and 6, compared with other strains, reduced the reduction of the ε-amine group the most. This result shows that the Lactobacillus or its fermented product of the present invention is helpful to improve the reduction of ε-amine content in the late stage of the anti-glycation reaction.
圖7係乳酸桿菌發酵物對蛋白質糖化後期產物游離硫氫基含量之影響。在對照組中，BSA (5 mg/mL) 與葡萄糖(10 mM)、MG (25 mM) 共同反應後，Glycated BSA 的游離硫氫基含量為0.97 nmoles/mg，標示如圖中偏中間側的垂直線。反觀加入乳酸桿菌發酵物的部分組別，游離硫氫基含量皆高於0.97 nmoles/mg，顯示乳酸桿菌發酵物還原被氧化的Glycated BSA之游離硫氫基。其中編號5和6的乳酸桿菌GKM3、GKK2的發酵物，相對於其他種菌株，其還原糖化蛋白硫氫基能力強。此結果說明本發明的乳酸桿菌或其發酵物，在抗糖化反應後期中具強還原硫氫基之能力。Fig. 7 shows the effect of lactobacillus fermentation product on the free sulfhydryl content of late product of protein saccharification. In the control group, after the co-reaction of BSA (5 mg/mL) with glucose (10 mM) and MG (25 mM), the free sulfhydryl content of Glycated BSA was 0.97 nmoles/mg, indicated by the middle side in the figure Vertical line. In contrast, in some groups added with Lactobacillus fermentation product, the free sulfhydryl content was higher than 0.97 nmoles/mg, indicating that the Lactobacillus fermentation product reduced the free sulfhydryl group of oxidized Glycated BSA. Among them, the fermentation products of Lactobacillus GKM3 and GKK2 numbered 5 and 6 have a stronger ability to reduce sulfhydryl groups of glycated proteins than other strains. This result shows that the lactobacillus or its fermented product of the present invention has a strong ability to reduce sulfhydryl groups in the late stage of the anti-glycation reaction.
實驗四：糖化後期高度糖化終產物Experiment 4: Highly glycosylated end products in the late stage of saccharification (AGEs)(AGEs) 含量的測定Determination of content
取用如實驗一中所配置的糖化反應液，在反應結束後第五天的反應液， 參考2013年Sero等人的方法，測反應MG衍生的高度糖化終產物 (AGEs) 含量。Taking the saccharification reaction solution as configured in
圖8係乳酸桿菌發酵物對蛋白質糖化後期高度糖化終產物 (AGEs)生成之影響。在一具體實施案例中，MG所衍生的典型AGEs產物之一為argpyrimidine。在對照組中，BSA (5 mg/mL) 與葡萄糖 (10 mM)、MG (25 mM) 共同反應後，argpyrimidine含量與未糖化前相比增加84.32倍，標示如圖中右側的垂直線(靠近圖8右側框線)。反觀加入乳酸桿菌發酵物組別，大部分argpyrimidine含量增加比例皆大幅度降低。此結果說明本發明乳酸桿菌或其發酵物，有助於減少糖化反應後期高度糖化終產物 (AGEs)的生成。Figure 8 shows the effect of lactobacillus fermentation on the formation of advanced glycation end products (AGEs) in the late stage of protein glycation. In a specific embodiment, one of the typical AGEs products derived from MG is argpyrimidine. In the control group, after the co-reaction of BSA (5 mg/mL) with glucose (10 mM) and MG (25 mM), the content of argpyrimidine increased 84.32 times compared with that before unsaccharification, indicated by the vertical line on the right side of the figure (near the frame line on the right side of Figure 8). On the other hand, when the lactobacillus fermented product was added, most of the argpyrimidine content increase ratios were greatly reduced. This result shows that the Lactobacillus or its fermented product of the present invention helps to reduce the generation of advanced glycated end products (AGEs) in the later stage of the saccharification reaction.
實驗五：糖化後期糖化蛋白與高度糖化終產物受器的結合能力的評估Experiment 5: Evaluation of the binding ability of glycated protein to receptors of highly glycated end products in the late stage of glycation
取用如實驗一中所配置的糖化反應液，在反應結束後第五天之反應液，參考1997年Du Yan等人的方法，以CircuLex AGE-RAGE in vitro Binding Assay Kit (No. CY-8151, MBL Medical & Biological Laboratories Co. Ltd.) 分析乳酸桿菌發酵物對MG-BSA 與 RAGE 結合之影響。Take the saccharification reaction solution as configured in
圖9係乳酸桿菌發酵物對糖化反應中高度糖化終產物 (AGEs)與高度糖化終產物受器(RAGE)結合之影響。因AGEs 結合細胞膜上之 RAGE 後會啟動特定細胞生物反應，相關反應於許多疾病及老化中扮演重要之角色。相較於以未添加乳酸桿菌發酵物的對照組之AGE-RAGE 結合率為100%作為基準，在乳酸桿菌發酵物加入的實驗組，其糖化過程中的AGE-RAGE 結合率皆下降。此結果說明本發明乳酸桿菌或其發酵物，有助於降低糖化反應中高度糖化終產物 (AGEs)與高度糖化終產物受器 (RAGE)之結合率。Figure 9 shows the effect of lactobacillus fermentation on the combination of advanced glycation end products (AGEs) and receptors for advanced glycation end products (RAGE) in the saccharification reaction. Because AGEs combine with RAGE on the cell membrane to initiate specific cellular biological responses, related responses play an important role in many diseases and aging. Compared with the AGE-RAGE binding rate of the control group without adding lactobacillus fermentation product as a benchmark, the AGE-RAGE binding rate during the saccharification process decreased in the experimental group added with lactobacillus fermentation product. This result shows that the Lactobacillus or its fermented product of the present invention helps to reduce the binding rate of advanced glycation end products (AGEs) and receptors for advanced glycation end products (RAGE) in the saccharification reaction.
實驗六：人體肌膚彈性度檢測實驗Experiment 6: Detection experiment of human skin elasticity
實驗設計受試者先以水份彈性檢測儀(ANENG品牌，昇旺電子企業社)分析前臂屈側之皮膚彈性。皮膚彈性分數的評估級距分為1到5分，個分數所分別表示的皮膚彈性狀況如下表：Experimental design The subjects first analyzed the skin elasticity on the flexion side of the forearm with a moisture elasticity tester (ANENG brand, Shengwang Electronics Co., Ltd.). The evaluation scale of the skin elasticity score is divided into 1 to 5 points, and the skin elasticity status represented by each score is shown in the following table:
表3
取得實驗前膚質之彈性數值後，每天塗抹定量100 μL的乳酸桿菌發酵液於前臂屈側之皮膚並記錄每天膚質彈性，實驗過程進行十天。After obtaining the skin elasticity value before the experiment, apply 100 μL of Lactobacillus fermented liquid on the skin of the flexion side of the forearm every day and record the daily skin elasticity. The experiment process lasted for ten days.
圖10係乳酸桿菌發酵物對人體肌膚彈性改善評估結果。經實測，乳酸桿菌發酵液對人體肌膚彈性度，隨著使用時間增長，膚質彈性也跟著上升。此結果說明本發明的乳酸桿菌或其發酵物，有助於增加肌膚彈性度，而能改善人體肌膚的老化現象。Figure 10 is the evaluation result of the improvement of human skin elasticity by the fermentation product of lactobacillus. According to the actual measurement, the elasticity of the lactobacillus fermented liquid on human skin increases with the increase of use time. This result shows that the lactobacillus or its fermented product of the present invention helps to increase skin elasticity and improve the aging phenomenon of human skin.
實驗七：人體肌膚水分檢測實驗Experiment 7: Human skin moisture detection experiment
實驗設計受試者先以水份彈性檢測儀(ANENG品牌，昇旺電子企業社)分析前臂屈側之皮膚水分。手臂水分的正常範圍的數值會隨著季節變換，而有所改變。夏季一般為35~60%，冬季則為夏季一般為30~45%。本實驗的檢測季節是冬季。取得實驗前肌膚水分數值後，塗抹定量100 μL乳酸桿菌發酵液於前臂屈側之皮膚並於十分鐘後再次紀錄肌膚水分。Experimental Design The subjects first analyzed the skin moisture on the flexion side of the forearm with a moisture elasticity tester (ANENG brand, Shengwang Electronics Co., Ltd.). The normal range of arm moisture will vary with the seasons. Summer is generally 35~60%, while winter is generally 30~45%. The detection season of this experiment is winter. After obtaining the skin moisture value before the experiment, apply 100 μL of Lactobacillus fermented liquid on the skin of the flexion side of the forearm and record the skin moisture again after 10 minutes.
圖11係乳酸桿菌發酵物對人體皮膚鎖水性結果。經實測，乳酸桿菌發酵液對人體肌膚鎖水性具提高的效果。說明本發明的乳酸桿菌或其發酵物有助於保持較好的肌膚彈性，而能改善人體肌膚老化現象之一的水分流失現象。Figure 11 is the result of the water-retaining properties of the lactobacillus fermented product on human skin. According to the actual measurement, the lactobacillus fermented liquid has the effect of improving the water-locking property of human skin. It shows that the lactobacillus or its fermented product of the present invention helps to maintain better skin elasticity, and can improve the water loss phenomenon, which is one of the aging phenomena of human skin.
無none
圖1顯示糖化反應及其產物的生成路徑與機制。Figure 1 shows the formation pathway and mechanism of the saccharification reaction and its products.
圖2顯示乳酸桿菌對糖化前期產物之影響。Figure 2 shows the effect of Lactobacillus on pre-glycation products.
圖3顯示乳酸桿菌對糖化中期雙羰基化合物的捕捉能力評估。Figure 3 shows the evaluation of the ability of Lactobacillus to capture dicarbonyl compounds in the mid-saccharification stage.
圖4顯示乳酸桿菌對糖化後期產物交聯反應之影響的電泳圖。Figure 4 shows the electropherogram showing the effect of Lactobacillus on the cross-linking reaction of late glycation products.
圖5顯示乳酸桿菌對糖化後期產物羰基含量之影響。Figure 5 shows the effect of Lactobacillus on the carbonyl content of late glycation products.
圖6顯示乳酸桿菌對糖化後期產物ε-胺基含量之影響。Figure 6 shows the effect of Lactobacillus on the content of ε-amine groups in late glycation products.
圖7顯示乳酸桿菌對糖化後期產物游離硫氫基含量之影響。Figure 7 shows the effect of Lactobacillus on the free sulfhydryl content of late saccharification products.
圖8顯示乳酸桿菌對糖化後期高度糖化終產物 （AGEs）之影響。Figure 8 shows the effect of Lactobacillus on advanced glycation end products (AGEs) in advanced glycation.
圖9顯示乳酸桿菌對糖化反應中高度糖化終產物 （AGEs）與高度糖化終產物受器（RAGE）結合之影響。Figure 9 shows the effect of Lactobacillus on the binding of advanced glycation end products (AGEs) and receptors for advanced glycation end products (RAGE) in the glycation reaction.
圖10顯示乳酸桿菌對人體肌膚彈性改評估。Figure 10 shows the evaluation of lactobacillus on the improvement of human skin elasticity.
圖11顯示乳酸桿菌對人體皮膚鎖水性結果。Figure 11 shows the results of the water retention of Lactobacillus on human skin.
胚芽乳酸桿菌（ Lactobacillus plantarum）GKM3：BCRC-910787 Lactobacillus plantarum GKM3: BCRC-910787
乳雙歧桿菌（ Bifidobacterium lactis）GKK2：BCRC-910826 Bifidobacterium lactis GKK2: BCRC-910826
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