WO2020073869A1 - 盐肤木果实或其提取物的用途 - Google Patents

盐肤木果实或其提取物的用途 Download PDF

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WO2020073869A1
WO2020073869A1 PCT/CN2019/109409 CN2019109409W WO2020073869A1 WO 2020073869 A1 WO2020073869 A1 WO 2020073869A1 CN 2019109409 W CN2019109409 W CN 2019109409W WO 2020073869 A1 WO2020073869 A1 WO 2020073869A1
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liver
extract
fruit
alcoholic fatty
group
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French (fr)
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蔡圣宝
吴子桓
王春丽
罗宗洪
陈态明
杨俊娴
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昆明理工大学
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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18Magnoliophyta (angiosperms)
    • A61K36/185Magnoliopsida (dicotyledons)
    • A61K36/22Anacardiaceae (Sumac family), e.g. smoketree, sumac or poison oak
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23LFOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23LFOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/105Plant extracts, their artificial duplicates or their derivatives
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/16Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23VINDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00Food compositions, function of food ingredients or processes for food or foodstuffs

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  • the present invention relates to the application of the salt skin fruit or its extract in the preparation of health food or medicine for treating and / or preventing alcoholic fatty liver and liver injury, and belongs to the technical field of food or medicine.
  • Alcoholic fatty liver disease is a hepatocellular injury disease caused by excessive drinking, which can be reversed or further developed into liver fibrosis and cirrhosis, manifested as progressive liver fatty degeneration, hepatitis, liver fiber And cirrhosis. Alcohol is slowly absorbed in the stomach in small amounts, mainly through simple diffusion in the duodenum and upper ileum. Only 2% to 10% are excreted through the lungs and kidneys, and the remaining 90% to 98% are oxidized and metabolized in the liver.
  • AFLD Alcoholic fatty liver disease
  • alcoholic fatty liver of which 10% to 35% of heavy drinkers will further develop alcoholic hepatitis, 5% to 15% Severe drinkers will develop cirrhosis.
  • AFLD accounts for a relatively high proportion of liver disease. In some countries where alcohol consumption is common, such as China, Russia, the United States, France, Germany, Italy, Spain, etc., the incidence of alcoholic liver disease is high. The data shows that the incidence of alcoholic liver cirrhosis It accounts for 50% to 75% of all liver cirrhosis.
  • Rhus chinensis Mill Fruit, also known as salt bran, traitor salt, salt plum, etc., is the fruit of Rhus chinensis. Born in the shrub and sparse forest of Lime Mountain at an altitude of 350-2300m. It is a common wild positive tree in China. Distributed in the northeast and Yunnan, Sichuan, Shanxi, Anhui, Jiangsu, Guangxi and other places. Rhus vulgaris does not have high requirements on soil conditions, and is commonly found in uncultivated land. It is also the main body of Chinese aphids gallnut. Salt skin wood has long been used to treat diseases in ancient China.
  • phloem leaf is used to treat inflammation and diarrhea; roots are used to treat malaria and jaundice; fruits and seeds are often used to treat dysentery.
  • the fruit can be used not only as an herbal medicine, but also as a condiment and beverage.
  • ripe fruits are often eaten fresh in some places in China.
  • the purpose of the present invention is to provide a new use of the salt skin fruit or its extract in medicine and health food, specifically to apply the salt skin fruit or its extract in the treatment and / or prevention of alcoholic fatty liver and liver New medicines and new health foods in injury-related diseases.
  • the invention is devoted to the experimental research on the prevention and treatment of alcoholic fatty liver and liver damage by the salt skin fruit, and it has been proved through multiple ways that the salt skin fruit or its extract has a significant protective effect on the alcoholic fatty liver and liver damage.
  • the extract of Phoebe bournei fruit is a water extract or an alcohol extract of Phoebe bournei fruit.
  • the present inventors conducted a series of studies on the prevention and treatment of alcoholic fatty liver and liver injury by Yanfumu fruit, and confirmed that the extract of Yanfumu fruit has a definite liver protection function, so it can be used for the prevention and treatment of alcoholic fatty liver and liver injury Preparation and development of new health food.
  • the extracts of syringa phyllostachys fruit used in the preparation of drugs and health foods for the prevention and treatment of alcoholic fatty liver and liver damage have the following advantages: the syringa phyllanthus plants can adapt to various harsh natural environments, and are widely distributed in China, and their fruit prices Very low-cost; the salt skin fruit in Yunnan and other places is often used as a raw material for the preparation of condiments and beverages. It is a traditional medicinal and edible fruit with high safety; the extraction and preparation process is simple and the market prospect is considerable , Contains huge social and economic effects.
  • the present invention has the following advantages:
  • the present invention has discovered new health foods or medical uses for the fruit of the salt skin, and opened up a new research field;
  • the present invention has found through experiments that different extracts of Pleurotus ostreatus fruit can effectively inhibit the deposition of fat particles in the liver of alcoholic fatty liver model rats at lower concentrations, and the extracts come from natural plants used as seasonings Raw materials, high safety.
  • Fig. 1 is the H & E staining diagram (H & E ⁇ 200) of liver pathological slices after intervention of different extracts of alcoholic fatty liver and sclerophyllum fruit caused by alcohol combined with high-fat feed in rats, where A is the control group; B is the model group ; C is the low concentration group of alcohol extract; D is the high concentration group of alcohol extract; E is the low concentration group of water extract; F is the high concentration group of water extract.
  • the model of alcoholic fatty liver was established by gavage alcohol combined with high-fat feed.
  • the animal model experiment was used to observe the preventive effect of the alcoholic extracts and water extracts of Pleurotus ostreatus fruit on alcoholic fatty liver and liver injury.
  • the extract of Phoebe bournei fruit mainly uses ethanol or boiling water to extract the active ingredients.
  • the specific method is as follows: the water extract mainly simulates the characteristics of torment in the pharmacopoeia, boiled in boiling water for 1 hour, the material-liquid ratio is 1:20; after boiling, it is centrifuged while hot, the speed is 4000r / min, and the supernatant is suction filtered Afterwards, the samples are concentrated and finally prepared by lyophilization and ready for use.
  • the alcohol extract is subjected to ultrasonic extraction using an ethanol solution with a mass concentration of 80%.
  • the extraction temperature is 25 ° C.
  • the material-liquid ratio is 1: 5
  • the extraction time is 30 min
  • the filter residue is extracted again, and then the supernatant is centrifuged at 4000 r / min to concentrate and freeze-dry be made of.
  • the male rats are all SPF grade SD rats 4-8 weeks old, and their body weight deviates by about 5% of their body weight at around 200g.
  • the experimental animal comes from Liaoning Changsheng Biotechnology Co., Ltd.
  • the rat number is SCKK (Liao) 2015-0001. Specifically, 60 mice are divided into 6 groups (10 mice per cage), with two groups of water extract and alcohol extract; the numbers are as follows:
  • Group 1 Control group (mainly gavage with distilled water, N), Group 2 model group (liquor gavage, M), Group 3 low concentration of alcohol extract 300mg / kg (alcohol and alcohol extract, AL), Group 4 Alcohol extract high concentration 900mg / kg (drinking wine and alcohol extract, AH), Group 5 water extract low concentration 300mg / kg (drinking wine and water extract, WL), Group 6 water extract high concentration 900mg / kg (Drinking wine and water extract, WH). All animals were fasted for 12 h before the first administration, and the weight of each mouse was recorded before administration (water is not prohibited). Rats were reared according to the standard rat rearing environment (after purchase, acclimated to this environment for seven days).
  • the method of making the model is as follows: every morning at 9 am with an intragastric administration volume of 5 mL / kg, the corresponding doses of two types of extracts or distilled water are infused, and then the corresponding volume of liquor is infused after 30 min (the control group does not inject liquor, use Distilled water instead, and the total volume of liquor and extract cannot exceed the volume of gavage). After gavage, each group was fed with high-fat feed. After 80 days of feeding, all rats were fasted for 12 hours, anesthetized rats with 10% chloral hydrate, weighed, and blood was taken to make serum for testing, followed by serum biochemical analysis; liver was weighed, and then taken The left lobe of the liver was immersed in 10% formalin solution and was later H & E stained.
  • the liver index of the high-fat model group was significantly increased (P ⁇ 0.05); while compared with the high-fat model group, the water extract and alcohol extract of the fruit of P. chinensis could significantly reduce the liver index ( P ⁇ 0.05), but there was no significant difference in the final body weight between the groups (P> 0.05).
  • the serum triglyceride and total cholesterol levels of the model group increased significantly (P ⁇ 0.05).
  • the water extract and the alcohol extract of P. chinensis fruit can significantly reduce the content of triglyceride and total cholesterol in serum (P ⁇ 0.05).
  • Pathological slices of rat liver are shown in Figure 1. Observation of alcoholic fatty liver slices under light microscope. The shape of liver cells in the blank group of rat slices is normal, the nucleus state is normal, and there is no obvious fat granule formation in liver cells. In the model group, the aggregation of nuclei and the damage of hepatocytes can be clearly observed. Not only that, there are also obvious fat particle formation in hepatocytes. In the slice of alcohol extract, the damage caused by alcohol to the liver is greatly reduced, and the formation of fat particles is very small. Similarly, the water extract of Phoebe bournei fruit can also significantly inhibit the damage of alcohol to liver cells and the formation of fat particles.

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Abstract

盐肤木果实或其提取物在制备治疗和/或预防酒精性脂肪肝及肝损伤的药物或保健食品中的用途。

Description

盐肤木果实或其提取物的用途 技术领域
本发明涉及盐肤木果实或其提取物在制备治疗和/或预防酒精性脂肪肝及肝损伤的保健食品或药品中的应用,属于食品或医药技术领域。
背景技术
酒精性脂肪肝(alcoholic fatty liver disease,AFLD)是过量饮酒引起的肝细胞损伤性疾病,可逆转也可进一步发展为肝纤维化和肝硬化,表现为渐进性的肝脏脂肪变性、肝炎、肝纤维化和肝硬化。酒精在胃中缓慢少量吸收,主要在十二指肠和上段回肠通过单纯扩散吸收。仅有2%~10%通过肺脏和肾脏排出,其余的90%~98%在肝脏内氧化代谢。大约90%的重度饮酒者(酒精摄入多于60克/天)患有酒精性脂肪肝,其中有10%~35%的重度饮酒者会进一步发展成酒精性肝炎,5%~15%的重度饮酒者会发展成肝硬化。AFLD在肝脏疾病发病中占相当高的比例,在一些饮酒普遍的国家如中国、俄罗斯、美国、法国、德国、意大利、西班牙等,酒精性肝病的发病率较高,资料显示酒精性肝硬化发病占所有肝硬化的50%~75%。饮酒是我国一个非常传统的饮食***不断增高,酒类产品的消耗量也逐年递增,从而导致人群中酒精性脂肪肝病的发病率明显上升。但是目前临床上对AFLD的治疗却缺乏有针对性的靶点治疗方法,只能被动的以对症处理为主(用营养支持来防治营养不良和用糖皮质激素来控制炎症),尚无防治酒精性脂肪肝及肝损伤的特效药。目前全世界都在积极寻找防治酒精性脂肪肝及肝损伤的方法及天然产物从而应对日益严峻的酒精性脂肪肝及肝损伤相关疾病问题。因此,对传统药食两用中草药防治酒精性脂肪肝及肝损伤的开发具有极其重要的意义。
盐肤木(Rhus chinensis Mill.)果实又名盐麸子、叛奴盐、盐梅子等,是漆树科植物盐肤木的果实。生于海拔350-2300m的石灰山灌丛、疏林中。为我国常见的野生阳性树。分布东北及云南、四川、山西、安徽、江苏、广西等地。漆树对土壤条件要求不高,常见于未耕地,也是众所周知的中国蚜虫五倍子的主体。盐肤木在中国古代各地早已用于治疗疾病。例如,盐肤木叶用于治疗炎症和腹泻;根用于治疗疟疾和黄疸;果实和种子经常用于治疗痢疾。与其他部位相比,果实不仅可以作为草药使用,还可以作为调味品和饮料使用。此外,成熟果实经常在中国的一些地方新鲜地吃。目前国内外对盐肤木果实提取物防治酒精性脂肪肝及肝损伤的相关研究未见有文献报道。
发明内容
本发明目的在于提供盐肤木果实或其提取物在医药和保健食品方面的新用途,具体而言就是将盐肤木果实或其提取物应用在制备治疗和/或预防酒精性脂肪肝及肝损伤等相关疾病中新药物和新保健食品中。
本发明致力于盐肤木果实防治酒精性脂肪肝及肝损伤的实验研究,经过多重途径证实盐肤木果实或其提取物对酒精性脂肪肝及肝损伤有显著的保护作用。
本发明中盐肤木果实提取物是盐肤木果实的水提取物或醇提取物。
本发明人对盐肤木果实防治酒精性脂肪肝及肝损伤进行了系列的研究,证实了盐肤木果实提取物具有确切的肝保护功能,因此可用于防治酒精性脂肪肝及肝损伤新药和新保健食品的制备与开发。盐肤木果实提取物用于制备防治酒精性脂肪肝及肝损伤药物和保健食品具有以下这些优势:盐肤木植物可以适应各种恶劣的自然环境,在我国的分布非常的广泛,其果实价格非常的低廉;在云南等地盐肤木果实经常被用作调味品和饮品的制作原料,是一种传统药食两用的果实,安全性高;提取和制备的工艺流程简单、市场前景可观、蕴藏着巨大的社会和经济效应。
与现有技术相比,本发明具有如下优点:
1、本发明为盐肤木果实发掘了新的保健食品或医疗用途,开拓了一个新的研究领域;
2、本发明通过实验发现盐肤木果实不同提取物在较低浓度下均可以有效的阻遏酒精性脂肪肝模型大鼠肝脏中脂肪颗粒的沉积,且提取物来自于用作调味料的天然植物原材料,安全性高。
附图说明
图1是酒精联合高脂饲料所致大鼠酒精性脂肪肝及盐肤木果实不同提取物干预后的肝脏病理切片的H&E染色图(H&E×200),其中A为对照组;B为模型组;C为醇提物低浓度组;D为醇提物高浓度组;E为水提物低浓度组;F为水提物高浓度组。
具体实施方式
下面结合病理切片附图和实验实例的数据来进一步的阐述本发明。这些实验的事例仅用于说明本发明,不用于限制本发明的应用范围。在阅读了本发明的记载以后,本领域的科技人员对其等效的各种改动、修改和修饰,都属于本发明权利要求所限定的范围。
酒精性脂肪肝的实例
应用灌胃酒精联合饲喂高脂饲料的方法建立酒精性脂肪肝的模型,通过动物模型实验观察盐肤木果实的醇提取物和水提取物对酒精性脂肪肝及肝损伤的防治作用。
1.实验材料
1.1动物、饲料及试剂
SD雄性大鼠,辽宁长生生物技术股份有限公司。高脂饲料,进多丰生物有限公司。56°红星二锅头,北京红星股份有限公司。试剂盒,南京建成生物科技有限公司。
1.2提取物
盐肤木果实提取物主要采用乙醇或沸水来提取其中有效成分。具体方法如下:水提取物主要模拟药典中煎熬的特性,在沸水中煮沸1小时,料液比为1:20;煮沸以后趁热离心,转速4000r/min,上清液进行抽滤,抽滤以后的样品进行浓缩,最后冻干制得,待用。
醇提取物采用质量浓度80%的乙醇溶液进行超声提取,提取温度为25℃,料液比为1:5提取时间为30min,滤渣复提一次,然后4000r/min离心取上清液浓缩冻干制得。
1.3仪器
全自动生化分析仪,深圳雷杜生命科学股份有限公司。
2.方法与结果
2.1造模
雄性大鼠都是4-8周龄的SPF级SD大鼠,体重在200g左右偏差5%左右的体重。实验动物来源于辽宁长生生物技术股份有限公司,大鼠的编号为SCKK(辽)2015-0001。具体是把60只老鼠分成6组(每笼10只),水提取物、醇提取物各两组;编号如下:
Group 1:对照组(主要以蒸馏水灌胃,N)、Group 2模型组(白酒灌胃,M)、Group 3醇提取物低浓度300mg/kg(灌酒和醇提取物,AL)、Group 4醇提取物高浓度900mg/kg(灌酒和醇提取物,AH)、Group 5水提取物低浓度300mg/kg(灌酒和水提取物,WL)、Group 6水提取物高浓度900mg/kg(灌酒和水提取物,WH)。第一次给药前所有动物禁食12h,并且在给药前记录各个老鼠的体重(不禁水)。按照标准的大鼠饲养环境饲养大鼠(购买后,在该环境下适应七天)。造模的方式如下:每天早上9点以5mL/kg的灌胃体积,灌入对应剂量的两类提取物或是蒸馏水,然后在30min以后灌入对应体积的白酒(对照组不灌白酒,用蒸馏水代替,且白酒和提取物的总共的体积不能超过灌胃的体积)。在灌胃以后,各个组给予高脂饲料的喂养。在饲养80天以后,对所有的大鼠禁食12h,使用10%的水合氯醛麻醉大鼠,称重,取血液制成血清待测,随后进行血清生化分析;取肝脏称重,随后取肝左叶,浸没在10%的***溶液中,待后续进行H&E染色。
2.2结果
2.2.1盐肤木果实提取物对酒精性脂肪肝大鼠体重和肝指数的影响
表1.盐肤木果实提取物对酒精性脂肪肝大鼠体重和肝指数的影响
Figure PCTCN2019109409-appb-000001
注: *表示模型组与空白对照组相比有显著性差异(P<0.05), #表示给药组与模型组相比有显著性差异(P<0.05);
与空白对照组相比,高脂模型组的肝指数显著的增加(P<0.05);而与高脂模型组相比,盐肤木果实的水提取物和醇提取物可以显著降低肝指数(P<0.05),但是最终体重各个组别之间没有显著的差异(P>0.05)。
2.2.2盐肤木果实提取物对酒精性脂肪肝大鼠血脂和肝功能酶的影响
表2.盐肤木果实提取物对酒精性脂肪肝大鼠血脂和肝功能酶的影响
Figure PCTCN2019109409-appb-000002
注: *表示模型组与空白对照组相比有显著性差异(P<0.05), #表示给药组与模型组相比有显著性差异(P<0.05);
与空白组相比,模型组血清中的甘油三脂和总胆固醇含量都显著性的上升(P<0.05)。而与模型组相比,盐肤木果实水提取物和醇提取物可以显著的降低血清中的甘油三脂和总胆固醇的含量(P<0.05)。另外,从表2中可以看出,与空白组相比,模型组血清中ALT和AST含量显著升高(P<0.05),说明酒精加高脂饮食对大鼠肝脏细胞造成了明显的损伤;但是与模型组相比,各给药组的大鼠血清中ALT和AST含量均显著下降(P<0.05),说明盐肤木果实提取物可以有效防治酒精造成的肝损伤,尤其是高浓度的醇提物,其对肝细胞酒精损伤的保护作用最强,ALT和AST基本恢复正常。
2.2.3盐肤木果实提取物对酒精性脂肪肝大鼠肝脂的影响
表3.盐肤木果实提取物对酒精性脂肪肝大鼠肝脂的影响
Figure PCTCN2019109409-appb-000003
注: *表示模型组与空白对照组相比有显著性差异(P<0.05), #表示给药组与模型组相比有显著性差异(P<0.05);
在动物的肝脏中,与空白对照组相比,高脂模型大鼠的肝中甘油三酯和总胆固醇的含量均显著升高(P<0.05)。而与高脂模型组相比,盐肤木果实的水提取物或醇提取物处理组的甘油三酯含量均显著下降(P<0.05),且与空白对照组没有显著差异(P>0.05),这一结果与对应的肝脏切片可以相互印证,进一步证实盐肤木果实的水或醇提取物可以有效 的防治酒精性脂肪肝。另外,高浓度醇提取物和低浓度水提取物可以显著降低肝脏中总胆固醇含量(P<0.05)。
2.2.4盐肤木果实提取物对酒精性脂肪肝大鼠肝组织病理情况的影响
大鼠肝脏病理切片如图1所示,在光镜下观察酒精性脂肪肝切片,空白组的大鼠切片中肝细胞的形状正常,细胞核状态正常,且肝细胞中没有明显的脂肪粒形成。模型组中,可以明显的观察到细胞核的聚集和肝细胞的损伤,不仅如此,在肝细胞中也有明显的脂肪粒形成。醇提取物的切片中酒精对肝造成的损伤大大的减少,且脂肪粒形成的很少。同样,盐肤木果实水提取物也可以显著地抑制酒精对肝细胞的损伤和脂肪粒的形成。
综上所述:可以确定盐肤木果实的提取物可以有效的抑制长期饮酒所造成的脂肪在肝细胞中的积累,且可以有效的阻止酒精对肝细胞的损伤。从而确定盐肤木果实及其提取物可以防治酒精性脂肪肝及肝损伤。

Claims (2)

  1. 盐肤木果实或其提取物在制备治疗和/或预防酒精性脂肪肝及肝损伤药物中的应用。
  2. 盐肤木果实或其提取物在制备治疗和/或预防酒精性脂肪肝及肝损伤保健食品中的应用。
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CN113134021A (zh) * 2021-04-13 2021-07-20 昆明理工大学 盐肤木果实或其提取物在制备防治药物性肝炎肝损伤药物中的应用
CN115054621B (zh) * 2022-01-03 2023-12-22 昆明理工大学 盐肤木果实或其提取物防治骨质疏松的用途
CN116602999A (zh) * 2022-02-08 2023-08-18 蔡圣宝 盐肤木果实提取物在制备黄嘌呤氧化酶抑制剂、防治高尿酸症药物、功能性食品中的应用
CN116603000A (zh) * 2022-02-09 2023-08-18 蔡圣宝 盐肤木果实提取物及其制备方法和在制备防治痛风的药物、功能性食品中的应用
CN116617280A (zh) * 2022-02-10 2023-08-22 蔡圣宝 盐肤木果实提取物在制备防治尿酸性肾病的药物、功能性食品中的应用

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