CN101361764B - Composition containing brown alga polysaccharide sulfuric ester and glossy ganoderma polysaccharide and use thereof - Google Patents
Composition containing brown alga polysaccharide sulfuric ester and glossy ganoderma polysaccharide and use thereof Download PDFInfo
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Abstract
The invention discloses a composition containing alga polysaccharides sulfuric acid ester and ganoderma lucidum polysaccharide. The invention also discloses an application of the composition in preventing and curing chemical liver injury and regulating immunity. The research of the inventor shows that the composition has good effect on regulating the balance of human immune system, enhancing resisted capacity against diseases and curing chemical liver injury if the composition is taken by patients for a long time. The invention also discloses a preparation technique of the composition.
Description
Technical field
The invention discloses a kind of compositions, disclosing a kind of especially is the Orally administered composition of active component with sulfated fucan and ganoderan class material, further, the invention discloses the application of said composition aspect the chemical hepatic injury of control, adjusting immunity.
Background technology
Chemical liver injury mainly comes from the harmful substance that working environment, food, medicine and heavy drinking bring, and these material types are contained inorganic toxic material (heavy metal, chloride etc.), pesticide, medicine and ethanol etc.As will the detoxify liver of organ of body weight for humans, have two channel blood supplies of Hepatic artery and hepatic vein, chemical substance can enter liver by gastrointestinal tract portal vein or body circulation and transform, so liver is subjected to the infringement of chemical toxicant in these materials easily.Since the industrial revolution, exist some to the virose material of liver in the Nature and the commercial production, research worker is referred to as " hepatotropic poison ", these materials easily cause ill in human body, incubation period is short; pathological process and inhalation dose or infective dose present positive correlation, can cause liver cell necrosis in different degree, fatty distortion, liver cirrhosis and hepatocarcinoma.Raw material in the commercial production, intermediate product and final products all may have liver toxicity, therefore need the generation of very vigilant chemical liver injury.This also requires research worker can study the medicine and the health product of some effectively preventing hepatic injury simultaneously.
Liver is the vitals of human body, and human liver disease patient is more and more at present, but still lacking curative effect up to now obviously has the little medicine of side effect concurrently.It is generally acknowledged that hepatocyte injury is the result of multiple effect.The pathomechanism of hepatic injury thinks relevant with the peroxidating of intrahepatic fat acid metabolic obstacle and free radical and initiation thereof more due to the chemical substance (such as chlorination carbon, ethanol etc.).
Research thinks that chemical toxicant may infect liver by following pathomechanism:
1. steatosis, carbon tetrachloride, yellow phosphorus etc. disturb the synthetic transhipment path of lipid protein, form fatty liver at last.
2. lipid peroxidation, this is a kind of specific form of expression of toxicity hepatic injury, for example carbon tetrachloride etc. produces the very strong intermediate product of a kind of oxidability in vivo during metabolism, the lipid that this intermediate product acts on the biological cell membrane forms peroxidating, thereby destroy the phospholipid layer space structure of film, cause the 26S Proteasome Structure and Function generation pathologic of cell to change.
3. cholestasis reaction, it is impaired mainly to come from liver cell film and microvillus, causes the bile acid acatharsia.
The hepatic injury that chemical substance causes can be caused by its substance metabolism in hepatocyte.Can suppress the mitochondrion tricarboxylic acid cycle by the value of rising NADH/NAD such as intracellular ethanol, cause intrahepatic fat acid metabolic obstacle, oxidation reduction and NADH increase, promoted the synthetic of fatty acid, make neutral fat in hepatocyte, pile up to take place steatosis, serious steatosis even cause proliferation of fibrous tissue in the hepatic necrosis regulating liver-QI.Liver damage disease, especially fatty liver and abnormal liver function follow blood lipid level to increase more, and this discloses, and fatty acid metabolism is disorderly to have certain relation with fatty liver and hepatic injury.
The lipid peroxidation that studies confirm that in recent years is an important chemical liver injury path, and body endoperoxides increased response can cause that antioxidant ability of organism descends, thereby the generation of free radical increases.Thing followed free radical is oxidation cell membrane lipid not only, the more important thing is can oxidation epicyte protein, thereby cause hepatocyte function and structure not normal.Free radical scavenger such as SOD and CAT can be removed the superoxide anion and the hydroxy radical that possess the height reactivity that produces in the lipid peroxidation respectively, the cell injury that the control lipid peroxidation causes.
Immune system is one of most important biosystem in the human body, and its function has a significant impact human body.In case the function of immune system imbalance occurs, will cause human body that disease body health ability is weakened.Immune disorder, immunologic hypofunction crowd ratio are increasing in recent years, will cause the disease of immune system patient sharply to increase for a long time in the past.Immune system is that human body is resisted pathogen and invaded most important safeguard system, this system is by immune organ (bone marrow, thymus, spleen, lymph node, tonsil, small intestinal aggregated lymphatic follicles, vermiform appendix etc.), immunocyte (lymphocyte, mononuclear phagocyte, neutrophilic granulocyte, basophil, eosinophilic granulocyte, mastocyte, platelet etc.), and immune molecule (complement, immunoglobulin, cytokine etc.) is formed.It is the most effective weapon of body defence pathogen invasion, but the hyperfunction meeting of its function is to self organ or tissue's generation injury.In a lot of self-diseases, the not normal important pathogenesis often of function of immune system.Each component function of immune system normally be to keep the metastable assurance of body's immunity, the hyperfunction of the defective of any component or function all can be brought infringement to body.
Each component of immune system blazons whole body, and is intricate, and particularly immunocyte and immune molecule constantly produce in body, circulate and upgrade.Immune system has the discriminative power of height, can accurately discern own and non-own material, to keep the relative stability of body; Can also accept, transmit, enlarge, store the information of relevant immunity simultaneously with memory, reply and constantly adjust its responsiveness at immunologic information generation positive and negative, therefore the imbalance of function of immune system also can be totally unfavorable to human body: the identification ability of human body is easy unusually to cause the generation of allergic phenomena (to use certain food, injectable drug anaphylaxis to occur, even cause suffering a shock), on the contrary then can cause repeated infection; The Selfstabilizing ability of human body is unusual, and immune system is reacted to the cell of self, causes autoimmune disease.This shows that the human immune system plays a part very important to human beings'health, if its functional lability, human body probably can be by virus, these pathogen infringements of antibacterial.
Sargassum is the important component part of marine living resources, and the Thallus Laminariae (Thallus Eckloniae) (Lam/nar/ajaponica aresch) of the brown sub-guiding principle Laminariales Laminariaceae Larminaria of Sargassum door is the highest Sargassum of output, and the Thallus Laminariae (Thallus Eckloniae) output of China occupies first place, the world.Thallus Laminariae (Thallus Eckloniae) claims the ocean vegetable again; contain the rich functions material; as trace element-selenium, cobalt, the chromium that iodine, mannitol, vitamin (A, B, C, F), taurine and needed by human body are wanted, be a kind of existing edibility, have again medicinal, the health care, industry, agronomical value the macro product.
Sulfated fucan is the Sulfated polysaccharide of a class, is present in the Brown algae, is called fucoidan (fucoidan or Brown algae polysaccharide, fucosan, sulfated fucan) according to international nomenclature principle unification.Now people have had comparatively clearly the composition of sulfated fucan and have understood, and it is a class chemical composition and the very complicated polysaccharide of structure, based on fucose and sulfate, along with the difference of algae also contains galactose, xylose, other compositions such as alduronic acid.Thallus Laminariae (Thallus Eckloniae) Fucoidan is by fucose.Galactose, xylose, alduronic acid, monosaccharide such as arabinose are formed.Based on fucose and galactose, the two ratio is approximately 3:1.
Existing research has confirmed that laminarin has good effect aspect control hepatic injury, the adjusting immunity.
Aspect the chemical hepatic injury that causes of the multiple reason of control, studies confirm that sulfated fucan can increase the activity of some antioxidase, reduce the generation of free radical, thus the attack that alleviates radical pair liver, renal tissue.Therefore, by removing free radical and antioxidation, sulfated fucan can be brought into play the liver protective effect.In addition, sulfated fucan can directly be removed ultra-oxygen anion free radical and hydroxyl radical free radical (0H) external, is a kind of good antioxidant; The ability that stronger removing reactive oxygen free radical is also arranged in vivo, the content of high lipid peroxide (LPO) significantly strengthens SOD activity in serum and the tissue in significantly reducing serum and organizing.
Aspect adjusting immunity, kelp polysaccharide sulfate is to influence (Liaocheng University's journal of Turnover of Mouse Peritoneal Macrophages activation and cytotoxicity, 2004.17 (2) 56-57) research thinks that kelp polysaccharide sulfate can activate Turnover of Mouse Peritoneal Macrophages, strengthen its immune function, opposing and kill tumor cell are had positive role.
It is that the laminarin of 15000D is to the cytophagous activation of mice that kelp polysaccharide sulfate has been studied mean molecule quantities such as a kind of L-of comprising fucose, D-galactose to the immunoregulation effect (Chinese immunology, 2000.2 (16)) of Turnover of Mouse Peritoneal Macrophages.Weigh with tumor control rate, suppress tumor cell pathological changes effective percentage and reached 86.5%.Effectively activating macrophage of laminarin is described, secretory cell toxic effect molecule improves macrophage peroxidase activity and phagocytic function.
Existing many pieces of documents disclose the preparation method of sulfated fucan.The clear 46-2248 of Japan Patent adopts hexadecane pyridinium chloride or hexadecane trimethyl ammonium bromide and sulfated fucan to be reacted into the quaternary amine complex, utilize the dissolubility difference of this complex again to salt, with ethanol, methanol and ion exchange resin treatment, purification is removed Algin, neutral polysaccharide and other impurity, and obtains the fucoidin sulphuric acid acid esters of comparison purification.CN1129109A then discloses the alkali condensation method by dried kelp soaking, filtration for several times, secondary ethanol extraction, washing with alcohol, fit adjustment PH scope etc.CN1344565A then discloses another kind of preparation method, comprises steps such as pretreatment of raw material, temperature control stirring and leaching, centrifugal, concentrated, ethanol precipitation, dehydrated alcohol dehydration.CN1517356A then is mixed with aqueous solution with fucoidan, adds hydrogen peroxide, hypochlorous acid or nitrous acid and salt thereof, and with the heating of gained mixed solution, the membrane ultrafiltration with molecular cut off 3000-5000 obtains fucoidan oligosaccharide.CN1616494A is a raw material with the natural seaweed sulfated polysaccharide, will add ascorbic acid and hydrogen peroxide, control reaction temperature in the Sargassum sulphuric acid sugar juice, the constant temperature degradation time is 0.5-3hr, dialysis or ultrafiltration again, concentrating under reduced pressure makes the low molecular weight seaweed polysaccharide sulfate product of 4-100KDa.In addition, CN1670028A, CN1392160A, CN1197674A also disclose methods such as adopting flocculation respectively and have prepared Sargassum polysaccharides.In the present invention, the technical scheme of above disclosure of the Invention all can be introduced into and obtain the used sulfated fucan of the present invention.
Ganoderma (Ganoderma Lucidum) is a Basidiomycetes, the Polyporaceae Ganoderma.On the botany meaning, Ganoderma comprises any in known nearly 200 kinds of Polyporaceae Ganoderma plants, and sense stricto Ganoderma is meant and is Pharmacopoeia of People's Republic of China (2005 versions, two ones) Ganoderma lucidum (Leyss. Ex Fr.) Karst. that is recorded or the dry sporophore of Ganoderma.Official description, Ganoderma can invigorating QI and tranquilization, flat coughing admittedly breathed heavily, and Ganoderma also is one of U.S.'s " medical herbs pharmacopeia " only ten kinds of Chinese herbal medicine of including simultaneously.Modern study shows that the active ingredient of Ganoderma mainly contains ganoderan, Ganodenic acid, adenosine, ergosterol, Saponin, purine, pyrimidine, alkaloid, lactone etc., ganoderan is the main component of Ganoderma pharmacologically active, one one of 05 edition Chinese Pharmacopoeia with ganoderan as the Ganoderma quality control standard.
Experiment shows that ganoderan plays main effect at aging, poisoning and oxygen-derived free radicals.But sample degeneration of M Φ film and necrosis that ganoderan inhibited oxidation agent tBOOH causes, the survival rate of raising cell, the mice oxygen free radical injury that alloxan is caused also has protective effect in addition.People such as Li Mingchun (ganoderan is to the influence of Turnover of Mouse Peritoneal Macrophages reactive oxygen free radical. Chinese J Pharmacol Toxicol, 2000,14 (1): 65-68) adopt the laser scanning co-focusing microscope technology, the equal one GL of dynamic monitoring ganoderan to the influence of Turnover of Mouse Peritoneal Macrophages (M Φ) reactive oxygen free radical content, experiment shows, ganoderan can reduce the generation of mouse peritoneal M Φ reactive oxygen free radical, and can antagonism the output of the mouse peritoneal M Φ reactive oxygen free radical that causes of PMA.
Existing research has also confirmed the application of ganoderan aspect adjusting immunity.Ganoderma spore polysaccharide is to immunoregulation effect (China Medicine University's journal of tumor-bearing mice, 2007.38 (2): 162-166), ganoderan is to influence (Ji'nan University's journal of mouse humoral immune function, 2003.24 (2): 51-53), ganoderan 151 clothes liquid to the research of immune function of mice influence (Taishan Hospital's journal, 2004.25 (6): 586-588) wait document adopt the kinds of experiments plan-validation ganoderan to regulating the effect of immunity.Chinese patent application CN02151784.3, CN03116403.X, CN200510049438.X etc. all disclose the application of ganoderan in regulating body immunity.
The production of ganoderan is outside the raw material divided by wild or tame Ganoderma sporophore, also can extract from the mycelium of Ganderma lucidum submerged fermentation or fermentation liquid.It is very big that the ganoderan molecular weight covers, and, scarcely is dissolved in alcohol in high concentration and is dissolved in hot water except the low molecule of fraction to hundreds thousand of from hundreds of, therefore suitable water extraction-alcohol precipitation technology relatively.
Summary of the invention
The object of the present invention is to provide a kind of compositions, preparation, the present invention simultaneously also discloses said composition, the preparation method of preparation, application.
The applicant discovers that by with sulfated fucan, ganoderan class combinations of substances control chemical liver injury, adjusting immunity are had collaborative, outstanding effect, and its life-time service safety and health-care effect all are better than existing product.Has possesses synergistic on the chemical liver injury of present composition control simultaneously, the adjusting immunity effect.Its curative effect is higher than separate constituent, has reduced the use amount of polysaccharide component generally.Security study finds that preparation of the present invention does not have tangible toxic reaction, is fit to take for a long time.
The present invention introduces sulfated fucan and ganoderan class material for the first time and makes up and be used for as the control chemical liver injury in this area.The present patent application people has determined feasibility that sulfated fucan and ganoderan class composition of matter are used to prevent and treat chemical liver injury first by lot of experiments.
The present invention introduces sulfated fucan for the first time in this area and the ganoderan compositions is used to regulate body immunity.The present patent application people has determined feasibility that sulfated fucan and ganoderan compositions are used to regulate body immunity first by lot of experiments.
Technical solution of the present invention provides the compositions of a kind of sulfated fucan and ganoderan, the ganoderan class material that comprises the active component sulfated fucan and have obvious synergistic activity, wherein ganoderan class material is selected from one or more of Ganoderma medical material, Ganoderma extract, Ganoderma spore powder extracts, Sporoderm-broken Ganoderma Lucidum Spore powder extract, Ganoderma mycelium extract etc., is preferably Ganoderma mycelium extract.When the present composition is used for pharmaceutical preparation, also optionally comprise pharmaceutically acceptable carrier.Accordingly, the invention provides the purposes that said composition is used to prevent and treat chemical liver injury, regulates immunity.
In technical scheme provided by the invention, the oral formulations of the present composition is provided, wherein contain the present composition and pharmaceutically acceptable carrier.Sulfated fucan and ganoderan class substance weight portion rate can be 1:100-100:1, are preferably 1:30-30:1, most preferably are 1:10-10:1.
The used sulfated fucan of the present invention seaweeds biology that has drawn from, comprise any plant that sulfated fucan can be provided, comprise Alga Sgrgassi Enerves, Thallus Laminariae, Sargassum fusiforme (Harv.) Setch, Mus tail algae, Thallus Sargassi Kjellmaniani, Thallus Laminariae (Thallus Eckloniae), yellow tang or Fucus Vesiculosus etc., preferred plant is a Thallus Laminariae (Thallus Eckloniae), no matter be artificial cultivation or spontaneous growth.The sulfated fucan method that obtains from these plants can be quoted the technical method described in the background technology, and the molecular weight that is obtained is on 100,000 order of magnitude levels.The sulfated fucan mean molecule quantity that obtains from Thallus Laminariae (Thallus Eckloniae), Alga Sgrgassi Enerves, Thallus Sargassi Kjellmaniani is respectively 350KD, 550KD, 980KD.When the present invention uses, can use the higher sulfated fucan of these molecular weight ratio, preferentially select for use molecular weight at the low-molecular-weight sulfated fucan of 5000D-200KD, used low-molecular-weight algal polysaccharide sulfate can extract from Sargassum also can be from the depolymerization of high molecular sulfated fucan.It is pointed out that lasting progress, can change for the sulfated fucan molecular weight that the present invention uses and drop in the scope of the invention along with technology.The purity of used sulfated fucan is not less than 50%, is preferably and is not less than 70%, more preferably is not less than 90%, and the best is to be not less than 95%.
According to used algal kind difference, there is certain difference in main group fucose, the sulfate content of the sulfated fucan that is obtained, but this species diversity does not influence the present invention is applied to prevent and treat chemical liver injury, the just reinforcement or the reduction of health-care effect that are brought, this belongs within the scope of the invention.The sulfated fucan of indication of the present invention contain chemically a class chemical substance of generally accepted sulfated fucan definition.
The used ganoderan class of the present invention material is not subjected to the restriction of extraction process.Can quote several different methods obtains.For example Guangzhou food industry science and technology (2003.19 (4): the existing technical method of 12-15) being mentioned that obtains ganoderan; Ganoderan biological function and Study on Extraction Method thereof (Heilungkiang medicine 2007.3 (20): the 207-209) technical method of being mentioned.
In the present invention, a preferred extraction process can be: use water extraction (preferentially with water-boiling method or water circumfluence method Ganoderma mycelium, water temperature 80-100 ℃, time 1-4 hour), then with alcohol precipitation (preferred alcohol, concentration 60-95%, temperature 4-30 ℃, time 3-12 hour), can obtain to be rich in the primary extract of ganoderan thus.Repeat said process 1-3 time.Get final product ganoderan of the present invention.
According to spirit of the present invention, those skilled in the art can learn that using some other medicinal herbs most in use uses the Ganoderma used with substituting the present invention to reach similar or identical implementation result.Functional component and document by the present chemical hepatic injury health product of some controls that gone on the market, for example Ganoderma Fructus Lycii compound anti-oxidation effect (food and machinery, 2004.6) discloses ganoderan, lycium barbarum polysaccharide all has good preventive and therapeutic effect to hydroxyl radical free radical, ultra-oxygen anion free radical etc.Those skilled in the art can know that lycium barbarum polysaccharide can play the effect of class such as ganoderan of the present invention equally.This contains within the scope of the present invention.
The inventor has studied simultaneously sulfated fucan ganoderan compositions has been added effect improved that vitamin C brought, applicant's experiment shows that vitamin C can further increase to effect of the present invention on the basis of the present invention's combination, therefore the present invention is also comprising such technical scheme: on the basis of the present composition, add appropriate vitamin C and improve wooden working of an invention effect.
The present composition can directly be made preparation, also can optionally add pharmaceutical carriers and make oral formulations then.The present composition has positive role to control anthropochemistry liver damage, adjusting immunity, and therefore going for sub-health population cooks health product.Health product general reference of the present invention can improve and protect the product of health state, comprises F﹠B.The food of indication includes but not limited to Semen Tritici aestivi goods, starch-based product, meat product, plant extract, milk breast, health promoting candy etc.; The beverage of indication comprises that the drinkable flowing product of everyone body comprises common beverage and the drinks that is used to improve specific function, such as sports drink.
The present invention is when being applied to health product, can also add other nutritional labelings and use when finally being prepared as product, these nutrient substance comprise starch, glucose, lactose, mannose, sucrose, syrup, Mel, several amino acids, calcium ferrum zinc, vitamin etc.
When the present composition is specifically used, can be used in combination with adjuvant.Adjuvant can comprise any pharmacy and last acceptable, the guaranteed adjuvants of safety of threpsology such as excipient, antioxidant, PH regulator, antiseptic, isotonic agent.
The present composition can be prepared as polytype, comprises capsule, tablet, powder, granule, oral liquid or syrup etc., is preferably capsule and oral liquid.The appearance existence form that changes product belongs to the technical scheme that those skilled in the art can associate, and the shape that therefore changes final products belongs to coverage of the present invention.
The applicant studies product function of the present invention, discovery is used separately with sulfated fucan or ganoderan and is compared, the present composition is used peroxide content, the interior antioxidant activity of raising body, removing free radical and the lipid peroxidation that can significantly reduce in liver, cerebral tissue and the blood, has collaborative curative effect.Take product of the present invention for a long time and have liver protection effect, and positive role is arranged preventing and/or treating chemical liver injury.The present invention that discovers of immunoregulation effect uses the effect that can significantly improve the adjusting body immunity, and can regulate a plurality of immune systems such as body fluid, lymph, cell simultaneously, has collaborative curative effect.
The specific embodiment
Following embodiment is intended to illustrate specific embodiments of the present invention, does not limit protection domain of the present invention.Used in an embodiment ganoderan is according to Instructions Page 6 of the present invention obtained.
Sulfated fucan 1mg
Ganoderan 10mg
Starch 18mg
Mannitol 18mg
Glucose 15mg
Microcrystalline Cellulose 10mg
Silica gel 2mg
The mentioned component stirring and evenly mixing is become compositions, make capsule behind the gravimetry.
Sulfated fucan 20mg
Ganoderan 5mg
Starch 20mg
Mannitol 22mg
Glucose 15mg
Microcrystalline Cellulose 10mg
Silica gel 2mg
The mentioned component stirring and evenly mixing is become compositions, make capsule behind the gravimetry.
Preparation embodiment 3
Sulfated fucan 50mg
Ganoderan 5mg
Starch 24mg
Mannitol 24mg
Glucose 15mg
Microcrystalline Cellulose 10mg
Silica gel 3mg
The mentioned component stirring and evenly mixing is become compositions, make capsule behind the gravimetry.
Sulfated fucan 40mg
Ganoderan 9mg
Heme iron 7mg
Vitamin C 8ug
Folic acid 400ug
Mentioned component mixing grind into powder is dissolved in deionized water to 1000ml, seals, sterilization and packing, be prepared as the oral liquid that meets specific standard.
The present invention prepares oral liquid and can amplify on consumption in large-scale production, fully carry out after the dissolving cannedly sealing then, sterilizing and be packed as oral liquid up to specification, and such as 500ml, 1L etc.
The comparative example 1
Sulfated fucan 20mg
Starch 20mg
Mannitol 24mg
Glucose 16mg
Microcrystalline Cellulose 10mg
Silica gel 2mg
The mentioned component stirring and evenly mixing is become compositions, make capsule behind the gravimetry.
The comparative example 2
Ganoderan 5mg
Starch 20mg
Mannitol 20mg
Glucose 16mg
Microcrystalline Cellulose 10mg
Silica gel 2mg
The mentioned component stirring and evenly mixing is become compositions, make capsule behind the gravimetry.
Experimental example 1
Experiment material: white mice is in a good state of health.The present composition, sulfated fucan: ganoderan 4:1, the water-soluble suspension that is made as.
Experimental project: 30 days feeding trials of acute toxicity test
Experimental technique:
1. acute toxicity test: select 40 of white mice, each 20 of male and female, body weight is 18-22g.Each is divided into four dosage groups 16.00,7.27,3.31 at random the male and female mice, and 1.50g/KG (bw) observes 15d, performance of record toxicity and death condition continuously behind the mouse stomach.
2.30d feeding trial: the same acute toxicity test of the sample of employing; selecting body weight for use is the Wistar rat of 55-70g; rat is divided into matched group and three dosage groups; high dose B group adopts 4.50g/kg (bw); middle dosage C group 0.90g/kg (bw), low dosage D organizes 0.16g/kg (bw), every group of ten rats; contrast A group feeding arm's length basis feedstuff, the feedstuff intake is calculated according to rat body weight 10%.Observed 30 days continuously.Observation index comprises:
(1) clinical observation: the general performance of animal, behavior, poisoning symptom, dead symptom, take by weighing body weight and intake weekly, calculate the food overall utilization.
(2) routine blood test and biochemical indicator: venule is got blood in test hentriaconta-sky, measures Hb H GB, erythrocyte RBC, leukocyte WBC.
1. the acute toxicity test result is as follows:
Table 1 present composition acute toxicity test result
The health product of supposing production contrast the highest matched group that we adopt with adult 60kg body weight daily intaking amount 0.05g/kg.bw, 320 times of the intake that is equivalent to be grown up.The experimental observation fortnight is not seen poisoning symptom, and death toll is zero.Tried thing to the anxious toxicity LD of the mice of two kinds of sexes
50All greater than 15.00g/kg.bw. according to existing toxicity grading, belong to nontoxic rank.
2.30 it feeding trial result
1. to the influence of rat body weight, the result is referring to accompanying drawing 1
Conclusion: each treated animal growth is normal.With matched group relatively, there is not significant difference (P〉0.05) with it in each dosage group
2. to the influence of rat food absorption, the result is referring to accompanying drawing 2
Conclusion: feeding animal 30 days, the refusing to eat phenomenon does not appear in animal.There are not significant difference (P〉0.05) in each dosage treated animal and control animals food absorption
3. (serum albumin unit is that g/dl, red blood cell count(RBC) are * 10 to the Serological testing result among this figure referring to accompanying drawing 3
12/ l) numeration of leukocyte is * 10
9/ l)
Conclusion: dosage group and matched group do not have significant significant difference (P〉0.05).
Comprehensive above-mentioned experimental result can think that this product has no side effect.
Experimental example 2
Materials and methods: Wistar rat, 188-212g; The present composition, specification are every part of 25mg, sulfated fucan: ganoderan 4:1, the water-soluble suspension that is made as; Carbon tetrachloride; Glutamate pyruvate transaminase ALT; Glutamic oxaloacetic transaminase, GOT AST; Triglyceride TG.Dosage grouping: three dosage group: 500mg/kd.bw.d (high dose group) are set, (250mg/kd.bw.d middle dosage group), (120mg/kd.bw.d low dose group), tried thing and be diluted to needed concentration, sulfated fucan matched group (being abbreviated as the A matched group), ganoderan matched group (B matched group) and carbon tetrachloride model group, negative control group are set in addition.The sulfated fucan matched group is given and sulfated fucan 250mg/kd.bw.d, and the ganoderan matched group is given and ganoderan 250mg/kg.bw.d, the carbon tetrachloride model group, and negative control group is given and distilled water.
Experimental technique: make acute chemical liver damage model with 2% carbon tetrachloride: each dosage group gives the present composition (irritating stomach amount 5.00ml/kg.bw) according to setting dosage, A, and the B matched group gives sulfated fucan, ganoderan respectively.Blank group, model group give distilled water, continuous 30 days.The 30th day with each treated animal fasting overnight 16 hours, then with model group, composition dosage group, A, the disposable filling stomach of B matched group gives carbon tetrachloride (2% concentration, 5ml/kg.Bw). dosage group, A, B matched group continue to tried to finish test behind the thing to 24 hour, put to death animal., get blood system from serum, measure TG content in ALT, AST and the hepatic tissue, and get liver and do the pathology section.It is technical specification with degeneration, necrosis etc. that histopathology is observed hepatocyte.ALT, AST, TG check result are referring to accompanying drawing 4,5,6.
As seen from the figure, the ALT of present composition high dose group is lower than carbon tetrachloride model group, significant difference; The AST of high dose group is lower than carbon tetrachloride model group, significant difference.It can also be seen that simultaneously under Isodose 250mg/kd.bw.d, effect of the present invention obviously is better than independent sulfated fucan, ganoderan, significant difference.Obtained the ALT index relatively respectively: 26.0%, 26.7%, the AST index: 7% improvement 8.3%,, can think that effect has obtained obvious progress.On the TG index, also have an appointment 5% improvement.
HCD structure observation result of science is referring to accompanying drawing 7
As seen from the figure, high dose group liver cell lesion degree of the present invention is significantly less than carbon tetrachloride model group, significant difference.On the pathological change degree, obviously be better than the independent application of sulfated fucan, ganoderan.The present composition has been obtained 11.9%, 12.0% improvement with respect to sulfated fucan, the independent effect of ganoderan under the Isodose, and it is obvious to improve effect.Above-mentioned result of the test shows, compositions disclosed in this invention can effectively be prevented and treated chemical hepatic injury, effectively be improved the human body immunity.The contrast matched group, it is obvious that the present composition is used to prevent and treat the chemical liver injury effect, and effect is significantly better than two independent components under the equal consumption.
Experimental example 3
Select female mice for use, be divided into four groups at random, body weight 18-22g.The present composition, wherein sulfated fucan: ganoderan 3:1 is dissolved in deionized water and makes suspension.(cyclophospha-mide Cy), is used for the hypoimmunity model construction to Cyclophosphamide for injection.Sheep red blood cell (SRBC) (sheep e-rythrocyte, SRBC).
According to the normal use amount adult of human body 60kg body weight daily intaking amount 0.05g/kg.Bw, relate to and be divided into high, normal, basic three dosage groups (dosage is equivalent to 5,2,1 times of normal person's use amount respectively), ten every group.Sulfated fucan matched group, ganoderan matched group, model group, matched group are set in addition, give sulfated fucan, ganoderan, distilled water, the distilled water of 2 times of normal human's consumptions respectively, be designated as A matched group, B matched group, model group respectively.Adopt gastric infusion, once a day.Measure every immune indexes after 20 days.Wherein in the time of the 15 day except that matched group the continuous 2d of every mouse peritoneal injection Cy50mg/kg (matched group waits the capacity normal saline), the animal model of preparation immunologic hypofunction. the every caudal vein of Ahau injects the india ink (0.1ml/10g) of centrifugal back 50%, get blood 40l from eyeball respectively behind 2min and the 10min, and it is added to 4ml0.1%NaHCO
3In the solution, the 600nm wavelength is measured absorbance, presses literature method and measures phagocytic index.Record mice body weight is lived and is got thymus extremely and claim weight in wet base, with mouse thymus weight/body weight (mg/g) as thymus index.The assay method of half hemolysis value, lymhocyte transformation rate adopts conventional literature method, such as document: the Huai Chinese yam polysaccharose Study immune regulation (time precious traditional Chinese medical science traditional Chinese medicines, 2007.18 the scheme DTH method of testing that (5)) is adopted is as follows: preparation DTH model, every Mus abdominal part was coated with 1%DNFB acetone Oleum Sesami solution 50ml sensitization in the 15 day, strengthen 1 time with dosage next day, prepare the immunocompromised model simultaneously except that matched group, this model building method is with reference to the above-mentioned model building method that is used to test thymus index, phagocytic index.After the sensitization three days, 1%DNFB solution evenly is applied in mouse right ear attacks.Attack 24h, crane one and put to death mice, cut left and right sides auricular concha, lay round auricle with diameter 9mm card punch and weigh, the difference of left and right sides auricle is the swelling degree.
The anti-fatigue ability test has nothing to do in immunity, does not therefore have model group, also need not to adopt the hypoimmunity model construction, only needs matched group to get final product.
The influence of 1 table 2 couple mice delayed allergy DTH as a result
As a result 2: the influence of table 3 pair normal mouse immune organ thymus index
As a result 3: the influence of table 4 pair mouse macrophage function
As a result 4: the influence of table 5 pair mouse lymphocyte conversion ratio
As a result 5: the influence of table 6 pair mouse humoral immune ability
As a result 6: the influence of table 7 pair mouse anti-reflecting fatigue ability
Contrast control experiment group data, the effect that the present invention improves immunity is better than using separately single component.The present invention regulates immunologic function has contained control agent liquid systems and phagocyte and lymphsystem, can omnibearing enhancing body immunity.Show: 1. omnibearing raise immunity under normal dose comprises humoral immunization, phagocyte, lymphocytic immunity.。2. compositions unit's concentration is improved doses (such as 5 times of amounts), can suppress human immunologic function's enhancing, perhaps reduce the human immunologic function.Illustrate that purposes of the present invention can the two-ways regulation immune function, to immunity is excited certain inhibitory action arranged under the high dose.
Claims (9)
1. a pharmaceutical composition contains sulfated fucan and ganoderan, and ganoderan wherein is selected from one or more of Ganoderma medical material, Ganoderma extract, Ganoderma spore powder extracts, Sporoderm-broken Ganoderma Lucidum Spore powder extract, Ganoderma mycelium extract.
2. according to the compositions of claim 1, wherein the weight ratio of sulfated fucan and ganoderan is 1: 100-100: 1.
3. according to the compositions of claim 1 or 2, it is the form of medicine or health product.
4. according to the compositions of claim 3, also contain pharmacy acceptable auxiliary or health product acceptable additive in the compositions.
5. according to the compositions of claim 1, it is characterized in that also containing in the compositions vitamin C.
6. according to claim 1,2 or 4 compositions, it is tablet, pill, capsule, granule, powder, oral liquid or syrup.
7. the purposes of the compositions of claim 1～6 in preparation hepatic or health product.
8. the compositions of claim 1～6 is in preparation treatment or the medicine of prevention chemical liver injury or the purposes in the health product.
9. the compositions of claim 1～6 is regulated the medicine of body immunity or the purposes in the health product in preparation.
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| CN102755440A (en) * | 2012-06-27 | 2012-10-31 | 大连龙康生物科技有限公司 | Medicinal composition for treating immunodeficiency |
| CN103271357A (en) * | 2013-05-31 | 2013-09-04 | 南京年吉冷冻食品有限公司 | Healthy flood and preparation method thereof |
| KR20150033910A (en) * | 2013-09-25 | 2015-04-02 | 주식회사 풀무원 | The spore of Ganoderma lucidum which has the weight loss effect and the dietary components using this spore of Garnoderma lucidum for weight loss |
| CN109953999B (en) * | 2019-04-08 | 2021-09-21 | 清华大学 | Composition comprising Ganoderma polysaccharide and Polyporus polysaccharide with immunity enhancing effect |
| CN111035649B (en) * | 2019-11-15 | 2021-03-19 | 臻元国际(香港)有限公司 | NMN + GLP compound nutritional supplement and preparation method and application thereof |
| CN111304091A (en) * | 2019-11-28 | 2020-06-19 | 昆明理工大学 | Method for improving ganoderic acid content in ganoderma lucidum cell culture |
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| CN1745641A (en) * | 2005-09-26 | 2006-03-15 | 全英滨 | Vegetative for tified anticancer flour |
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