CN111297887B - Preparation method and application of liver-protecting active component of Yunnan ginseng - Google Patents
Preparation method and application of liver-protecting active component of Yunnan ginseng Download PDFInfo
- Publication number
- CN111297887B CN111297887B CN202010078020.6A CN202010078020A CN111297887B CN 111297887 B CN111297887 B CN 111297887B CN 202010078020 A CN202010078020 A CN 202010078020A CN 111297887 B CN111297887 B CN 111297887B
- Authority
- CN
- China
- Prior art keywords
- polysaccharide
- ginseng
- liver
- white
- liver injury
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 241000208340 Araliaceae Species 0.000 title claims abstract description 44
- 235000005035 Panax pseudoginseng ssp. pseudoginseng Nutrition 0.000 title claims abstract description 44
- 235000003140 Panax quinquefolius Nutrition 0.000 title claims abstract description 44
- 235000008434 ginseng Nutrition 0.000 title claims abstract description 44
- 238000002360 preparation method Methods 0.000 title claims abstract description 10
- 229920001282 polysaccharide Polymers 0.000 claims abstract description 67
- 239000005017 polysaccharide Substances 0.000 claims abstract description 67
- 150000004676 glycans Chemical class 0.000 claims abstract description 65
- 206010067125 Liver injury Diseases 0.000 claims abstract description 27
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims abstract description 21
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 16
- 230000000694 effects Effects 0.000 claims abstract description 13
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 10
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 9
- 238000001556 precipitation Methods 0.000 claims abstract description 4
- 238000005238 degreasing Methods 0.000 claims abstract 2
- 231100000753 hepatic injury Toxicity 0.000 claims description 23
- 238000000034 method Methods 0.000 claims description 15
- 210000002966 serum Anatomy 0.000 claims description 13
- 239000003814 drug Substances 0.000 claims description 12
- 230000001154 acute effect Effects 0.000 claims description 11
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 11
- 239000002244 precipitate Substances 0.000 claims description 10
- VYQNWZOUAUKGHI-UHFFFAOYSA-N monobenzone Chemical compound C1=CC(O)=CC=C1OCC1=CC=CC=C1 VYQNWZOUAUKGHI-UHFFFAOYSA-N 0.000 claims description 7
- 230000002757 inflammatory effect Effects 0.000 claims description 6
- 230000001376 precipitating effect Effects 0.000 claims description 6
- 238000010298 pulverizing process Methods 0.000 claims description 6
- 239000006228 supernatant Substances 0.000 claims description 5
- 238000002156 mixing Methods 0.000 claims description 4
- 108090001005 Interleukin-6 Proteins 0.000 claims 1
- 230000036542 oxidative stress Effects 0.000 abstract description 6
- 230000003908 liver function Effects 0.000 abstract description 5
- 239000000463 material Substances 0.000 abstract description 5
- 231100000439 acute liver injury Toxicity 0.000 abstract description 3
- 231100000331 toxic Toxicity 0.000 abstract description 3
- 230000002588 toxic effect Effects 0.000 abstract description 3
- 241000357625 Campanumoea Species 0.000 abstract 1
- 230000000857 drug effect Effects 0.000 abstract 1
- 210000004185 liver Anatomy 0.000 description 12
- 241000965254 Apostichopus japonicus Species 0.000 description 11
- 102100036475 Alanine aminotransferase 1 Human genes 0.000 description 10
- 108010082126 Alanine transaminase Proteins 0.000 description 10
- 108010003415 Aspartate Aminotransferases Proteins 0.000 description 10
- 102000004625 Aspartate Aminotransferases Human genes 0.000 description 10
- 241000699666 Mus <mouse, genus> Species 0.000 description 8
- 235000018102 proteins Nutrition 0.000 description 8
- 229940079593 drug Drugs 0.000 description 7
- 239000000243 solution Substances 0.000 description 6
- 241000699670 Mus sp. Species 0.000 description 4
- 201000010099 disease Diseases 0.000 description 4
- 210000005228 liver tissue Anatomy 0.000 description 4
- 238000002835 absorbance Methods 0.000 description 3
- 239000004480 active ingredient Substances 0.000 description 3
- 208000019423 liver disease Diseases 0.000 description 3
- OQUKIQWCVTZJAF-UHFFFAOYSA-N phenol;sulfuric acid Chemical compound OS(O)(=O)=O.OC1=CC=CC=C1 OQUKIQWCVTZJAF-UHFFFAOYSA-N 0.000 description 3
- 206010003827 Autoimmune hepatitis Diseases 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- -1 alba polysaccharide Chemical class 0.000 description 2
- 230000001363 autoimmune Effects 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 239000008187 granular material Substances 0.000 description 2
- 239000007902 hard capsule Substances 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 230000001900 immune effect Effects 0.000 description 2
- JMZOMFYRADAWOG-UHFFFAOYSA-N methyl 7-methoxy-4-(7-methoxy-5-methoxycarbonyl-1,3-benzodioxol-4-yl)-1,3-benzodioxole-5-carboxylate Chemical compound COC(=O)C1=CC(OC)=C2OCOC2=C1C1=C2OCOC2=C(OC)C=C1C(=O)OC JMZOMFYRADAWOG-UHFFFAOYSA-N 0.000 description 2
- 210000000056 organ Anatomy 0.000 description 2
- 239000008194 pharmaceutical composition Substances 0.000 description 2
- 230000000144 pharmacologic effect Effects 0.000 description 2
- 239000006187 pill Substances 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 239000007901 soft capsule Substances 0.000 description 2
- 239000003826 tablet Substances 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 206010067484 Adverse reaction Diseases 0.000 description 1
- 208000023275 Autoimmune disease Diseases 0.000 description 1
- 235000014145 Caesalpinia bonduc Nutrition 0.000 description 1
- 244000036978 Caesalpinia bonduc Species 0.000 description 1
- 235000016513 Caesalpinia crista Nutrition 0.000 description 1
- 108090000695 Cytokines Proteins 0.000 description 1
- 102000004127 Cytokines Human genes 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 206010019663 Hepatic failure Diseases 0.000 description 1
- 206010019668 Hepatic fibrosis Diseases 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 102100037850 Interferon gamma Human genes 0.000 description 1
- 108010074328 Interferon-gamma Proteins 0.000 description 1
- 102000004856 Lectins Human genes 0.000 description 1
- 108090001090 Lectins Proteins 0.000 description 1
- 210000001744 T-lymphocyte Anatomy 0.000 description 1
- 208000038016 acute inflammation Diseases 0.000 description 1
- 230000006022 acute inflammation Effects 0.000 description 1
- 230000006838 adverse reaction Effects 0.000 description 1
- 230000003064 anti-oxidating effect Effects 0.000 description 1
- 230000000740 bleeding effect Effects 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 210000005252 bulbus oculi Anatomy 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 230000005779 cell damage Effects 0.000 description 1
- 208000037887 cell injury Diseases 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 208000037976 chronic inflammation Diseases 0.000 description 1
- 230000006020 chronic inflammation Effects 0.000 description 1
- 231100000012 chronic liver injury Toxicity 0.000 description 1
- 208000019425 cirrhosis of liver Diseases 0.000 description 1
- 230000017119 detection of oxidative stress Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 239000003937 drug carrier Substances 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 235000019197 fats Nutrition 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 238000003304 gavage Methods 0.000 description 1
- 239000003862 glucocorticoid Substances 0.000 description 1
- 210000003494 hepatocyte Anatomy 0.000 description 1
- 241000411851 herbal medicine Species 0.000 description 1
- 230000007365 immunoregulation Effects 0.000 description 1
- 230000008595 infiltration Effects 0.000 description 1
- 238000001764 infiltration Methods 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 239000002523 lectin Substances 0.000 description 1
- 210000005229 liver cell Anatomy 0.000 description 1
- 231100000835 liver failure Toxicity 0.000 description 1
- 208000007903 liver failure Diseases 0.000 description 1
- 239000002445 liver protective agent Substances 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 210000004698 lymphocyte Anatomy 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 230000002297 mitogenic effect Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000017074 necrotic cell death Effects 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 210000004738 parenchymal cell Anatomy 0.000 description 1
- 230000008506 pathogenesis Effects 0.000 description 1
- 230000036285 pathological change Effects 0.000 description 1
- 231100000915 pathological change Toxicity 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000003449 preventive effect Effects 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000007670 refining Methods 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 230000011506 response to oxidative stress Effects 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 239000003440 toxic substance Substances 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, 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
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/125—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives containing carbohydrate syrups; containing sugars; containing sugar alcohols; containing starch hydrolysates
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/25—Araliaceae (Ginseng family), e.g. ivy, aralia, schefflera or tetrapanax
- A61K36/258—Panax (ginseng)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/34—Campanulaceae (Bellflower family)
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Molecular Biology (AREA)
- Engineering & Computer Science (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Veterinary Medicine (AREA)
- Organic Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Polymers & Plastics (AREA)
- Animal Behavior & Ethology (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Epidemiology (AREA)
- Sustainable Development (AREA)
- Biochemistry (AREA)
- Materials Engineering (AREA)
- Gastroenterology & Hepatology (AREA)
- Mycology (AREA)
- Nutrition Science (AREA)
- Food Science & Technology (AREA)
- Medicines Containing Plant Substances (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention belongs to the field of biotechnology, and discloses white cloud ginseng (Campanumoea javanicaBI.) polysaccharide preparation, protection against acute liver injury and application thereof. Degreasing the Yunnan ginseng medicinal material by using methanol, extracting the degreased Yunnan ginseng medicinal material by water at 80 ℃, and carrying out alcohol precipitation and protein removal on the obtained extract to finally obtain the polysaccharide of the Yunnan ginseng. The white-cloud ginseng polysaccharide can obviously improve liver function indexes and oxidative stress indexes, has the characteristics of small toxic and side effects and obvious drug effect, and has wide application prospects.
Description
Technical Field
The invention belongs to the technical field of biology, and particularly relates to an extraction method of white stichopus japonicus polysaccharide and application of the white stichopus japonicus polysaccharide in preventing and treating acute liver injury diseases.
Background
The liver is the largest parenchymal organ of the human body, not only participates in the metabolism of vital substances such as saccharides, fat, protein, vitamins and the like, but also is an important organ for detoxifying the human body, and is one of the most easily attacked main organs of toxic substances. Although there are many effective preventive and therapeutic means and drug treatment methods for liver diseases, there are still many disadvantages. The liver diseases have become diseases with extremely high mortality rate worldwide due to the problems of differences of autoimmune systems of patients, inadvisable attention of patients in early stage of diseases, low compliance and the like.
Autoimmune hepatitis is a common clinical disease type, and is an autoimmune disease mainly expressed by acute and chronic inflammation of the liver, lymphocyte infiltration and hepatocyte necrosis. With the progress of the course of disease, acute and chronic liver injury may develop into refractory liver diseases such as liver failure, hepatic fibrosis and liver cirrhosis if not treated effectively in time. The existing clinical treatment medicines mainly for treating autoimmune liver injury mainly comprise glucocorticoid and other medicines, and have great adverse reactions.
In the search for a liver-protecting drug with low toxicity and remarkable effect, people aim at nontoxic polysaccharide macromolecular organisms. The white Yunnan ginseng is a plant with extremely high polysaccharide content, and the medicinal material is one of Chinese herbal medicines commonly used in Yi medicine and Miao medicine of Yunnan province. The active ingredient of the polysaccharide has pharmacological effects of immunoregulation, antioxidation and the like.
Canavalin a (ConA) is a strong mitogenic lectin. The ConA-induced acute immune liver injury model is a classic auxiliary T cell-mediated liver cell injury model and can better simulate the pathogenesis and pathological changes of autoimmune hepatitis. The research establishes a ConA-induced mouse acute immunological liver injury model, and evaluates the liver protection efficacy of the white-clouded ginseng polysaccharide by using the liver function index, the serum cytokine and the oxidative stress index of the mouse after the white-clouded ginseng polysaccharide is dried.
Disclosure of Invention
The invention aims to provide a preparation method and application of a liver-protecting active component of white cloud ginseng.
In order to achieve the purpose of the invention, the invention provides the following technical scheme:
application of radix Ginseng Indici polysaccharide in preparing medicine for preventing or treating liver injury disease is provided.
The preparation method of the white cloud ginseng polysaccharide comprises the following steps: pulverizing radix Gentianae, reflux-defatting with methanol, extracting with water at 80 deg.C, precipitating with 80% ethanol to obtain crude polysaccharide, removing protein by sevege method, and discarding precipitate to obtain refined polysaccharide. The method comprises the following specific steps:
1. pulverizing radix Ginseng alba, and defatting with methanol at 60 deg.C.
2. Extracting defatted radix Ginseng alba polysaccharide with water at 80 deg.C for 3 hr for 2 times, mixing extractive solutions, and concentrating under reduced pressure.
3. Precipitating the concentrated extractive solution with 80% ethanol, centrifuging, and removing supernatant.
4. Removing protein from the crude polysaccharide after alcohol precipitation by a sevege method, centrifuging, and removing the precipitate for 5 times till no precipitate exists.
5. And detecting the content of the polysaccharide of the white stichopus japonicus after protein removal by a sulfuric acid phenol method.
The polysaccharide can be used for preventing or treating liver injury by reducing alanine Aminotransferase (ALT) and aspartate Aminotransferase (AST) in blood serum.
The polysaccharide can be used for preventing or treating liver injury by reducing serum inflammatory factor (IL-6) value.
The radix Caesalpiniae Ministry polysaccharide can prevent or treat liver injury diseases by increasing SOD value and MDA value of medium oxidative stress index of liver and reducing GSH-PX value.
The dose of the polysaccharide is 2-3 times per day, and 0.8-3.6g per time.
The dose of the polysaccharide is 2g for 3 times per day.
The preparation of the white cloud ginseng polysaccharide is characterized in that the white cloud ginseng polysaccharide is added with pharmaceutically acceptable auxiliary materials to be prepared into tablets, hard capsules, soft capsules, powder, pills and granules.
A pharmaceutical composition comprises polysaccharide of radix Ginseng alba as active ingredient or pharmaceutically acceptable carrier.
In order to confirm the liver protection activity of the white cloud ginseng polysaccharide, the invention adopts a ConA induced mouse acute immune liver injury model to evaluate the prevention and treatment effect of the white cloud ginseng polysaccharide on liver injury, and the experimental result shows that: the polysaccharide of the white cloud ginseng can obviously reduce liver function indexes, serum inflammatory factors and improve oxidative stress indexes.
The invention has the advantages that:
1. the preparation method of the invention uses the white cloud ginseng as the raw material, and the polysaccharide component with higher content can be effectively and quickly obtained.
2. Pharmacological activity experiments prove that the polysaccharide of the white stichopus japonicus has the liver protection effect, has the characteristic of small toxic and side effects, and can be used for research and development of liver protection health care products and medicines.
Drawings
In order to make the purpose, technical scheme and experimental result of the invention clearer, the invention is illustrated by the following drawings:
in all the figures, NC is a normal control group, MC is a model control group, PC is a bifendate positive control group, BL is a white ginseng low-dosage group of 400mg/kg, BH is a white ginseng high-dosage group of 1200mg/kg; in comparison with the NC group, "#" indicates P <0.05, "# #" indicates P < 0.01, "# # ##" indicates P <0.001; in comparison to the MC group, "×" indicates P <0.05, "×" indicates P < 0.01, "×" indicates P <0.001;
FIG. 1 shows the intervention effect of Caesalpinia crista polysaccharide on liver function index (a: AST, b: ALT) of ConA-induced acute immune liver injury model in mice;
FIG. 2 shows the effect of the polysaccharide on the ConA-induced serum inflammatory factor (IL-6) in the mouse acute immune liver injury model;
FIG. 3 shows the effect of the polysaccharide on the oxidative stress index (a: SOD, b: MDA, c: GSH-PX) in ConA-induced mouse acute immune liver injury model.
Detailed Description
The technical solutions of the present invention are further described in detail below with reference to the examples, but the present invention is not limited in any way, and any modifications based on the present invention are within the scope of the present invention.
Application of radix Ginseng Indici polysaccharide in preparing medicine for preventing or treating liver injury disease is provided.
The preparation method of the white cloud ginseng polysaccharide comprises the following steps: pulverizing radix Gentianae, reflux-defatting with methanol, extracting with water at 80 deg.C, precipitating with 80% ethanol to obtain crude polysaccharide, deproteinizing by sevege method, and removing precipitate to obtain refined polysaccharide. The method comprises the following specific steps:
1. pulverizing radix Ginseng alba, and defatting with methanol at 60 deg.C.
2. Extracting defatted radix Ginseng alba polysaccharide with water at 80 deg.C for 3 hr for 2 times, mixing extractive solutions, and concentrating under reduced pressure.
3. Precipitating the concentrated extractive solution with 80% ethanol, centrifuging, and removing supernatant.
4. Removing protein from the crude polysaccharide after alcohol precipitation by a sevege method, centrifuging, and discarding the precipitate for 5 times until no precipitate exists.
5. And detecting the content of the polysaccharide of the white stichopus japonicus after protein removal by a sulfuric acid phenol method.
The radix Caesalpiniae Ministrae polysaccharide can prevent or treat liver injury diseases by reducing alanine Aminotransferase (ALT) value and aspartate Aminotransferase (AST) value in blood serum.
The polysaccharide can be used for preventing or treating liver injury by reducing serum inflammatory factor (IL-6).
The polysaccharide can be used for preventing or treating liver injury by increasing SOD value and MDA value of medium oxidative stress index of liver, and reducing GSH-PX value.
The dose of the polysaccharide is 2-3 times per day, and 0.8-3.6g per time.
The dose of the polysaccharide is 2g for 3 times per day.
The preparation of the white cloud ginseng polysaccharide is characterized in that the white cloud ginseng polysaccharide is added with pharmaceutically acceptable auxiliary materials to be prepared into tablets, hard capsules, soft capsules, powder, pills and granules.
A pharmaceutical composition contains polysaccharide of radix Ginseng alba as active ingredient or medicinal carrier.
Example 1
Preparing the crude polysaccharide of the white Yunnan ginseng:
pulverizing radix Ginseng Indici, and defatting with methanol at 60 deg.C for 3 times. Decocting the defatted radix Ginseng alba with hot water at 80 deg.C for 3 hr, extracting twice, mixing extractive solutions, and concentrating under reduced pressure. Precipitating the concentrated extract with 80% ethanol, centrifuging, and removing supernatant to obtain crude polysaccharide of radix Ginseng alba.
Example 2
Refining the white cloud ginseng polysaccharide:
redissolving the obtained polysaccharide of the white stichopus japonicus with distilled water, removing protein by a sevege method, centrifuging, discarding the precipitate for 5 times totally until no precipitate is generated after centrifuging. And finally, decompressing and concentrating the polysaccharide after protein removal, freezing and drying, and detecting the content of the polysaccharide by a sulfuric acid-phenol method. The content of the finally obtained white stichopus japonicus polysaccharide is 96.48 percent.
Example 3
Liver protection efficacy research of white cloud ginseng polysaccharide
Cloudburst ginseng polysaccharide is provided by example 2.
1. Animal grouping, modeling and administration method
ICR mice were randomly divided into a normal control group, a model group, a positive group (bifendate 150 mg/kg), a low-dose group (400 mg/kg) of white-cloud ginseng polysaccharide and a high-dose group (1200 mg/kg) of white-cloud ginseng polysaccharide, each group containing 10 mice. The administration was performed by gavage, and the normal group and the model group were administered with physiological saline, and the other groups were administered with the corresponding dose of the drug 1 time for 1 day for 7 consecutive days. After 30min of the last administration, 12mg/kgConA is intravenously injected to the model group and each administration group to prepare an acute immune liver injury model, and normal groups are intravenously injected with normal saline with the same volume. After fasting for 7h without water deprivation, the animals were sacrificed by orbital bleeding and liver collection.
2. Serum liver function index detection
Blood is taken from eyeballs in a 1.5mL centrifuge tube, and is centrifuged at 4500r/min at normal temperature for 10min, and supernatant is left. Processing according to the procedures of alanine Aminotransferase (ALT) and aspartate Aminotransferase (AST) kits, and measuring the absorbance values (OD values) of ALT and AST in serum by using a microplate reader. And calculating the corresponding enzyme activity value according to the OD value by searching a standard curve.
The experimental results are shown in fig. 1, compared with the normal group, the AST and ALT values of the model group are obviously increased, and compared with the model group, the AST and ALT values of the positive drug and the administration groups with different doses are obviously reduced, and the dose dependence is shown. The result shows that the polysaccharide of the white stichopus japonicus has a protective effect on mice with acute immunological liver injury.
3. Detection of inflammatory factor indicators in serum
And (3) detecting IL-6 and IFN-gamma in the serum by an ELISA method. Processing according to the IL-6 kit operation steps, measuring the IL-6 absorbance (OD) value in serum by using a microplate reader, and calculating by searching a standard curve.
The experimental result is shown in figure 2, compared with the normal group, the IL-6 of the model group is obviously increased, compared with the model group, the administration group with different dosage is obviously reduced, and the white stichopus japonicus polysaccharide can reduce the inflammatory reaction of the liver injury mouse.
4. Detection of oxidative stress indicators in liver tissue
Homogenizing mouse liver tissues, and according to the weight of the liver tissues: saline = 1.9 a 10% tissue homogenate was prepared for testing. Treating according to the operation steps of the SOD, MDA and GSH-PX reagent kit, measuring the absorbance values of the SOD, MDA and GSH-PX in the liver tissue by using an enzyme labeling instrument, and calculating by using a corresponding formula.
The experimental result is shown in figure 3, compared with the normal group, the SOD and the GSH-PX of the model group are obviously reduced, and compared with the model group, the SOD and the GSH-PX of each administration group are obviously increased; compared with the normal group, the MDA of the model group is obviously increased, and compared with the model group, the MDA of each administration group is obviously reduced, and the result shows that the polysaccharide of the white stichopus japonicus can improve the oxidative stress reaction caused by liver injury.
In conclusion, the white cloud ginseng polysaccharide has the characteristics of obvious liver protection activity and small toxic and side effects, and has wide application in the development of liver protection health care products and medicines.
In the mouse acute immune liver injury test, the dosage of the white stichopus japonicus polysaccharide is 400-1200mg/kg/d. Converted into human (calculated by adult weight 60 kg) dosage of 2.4-7.2g/d. In consideration of the difference in body weight of human and various other pathological conditions, the recommended dose is 2-3 times daily, 0.8-3.6g each time. The most common dose is 2g each time 3 times daily.
Claims (1)
1. The application of the white cloud ginseng polysaccharide in preparing the medicine for preventing or treating the acute immune liver injury diseases achieves the effect of preventing or treating the liver injury diseases by reducing the IL-6 value of a serum inflammatory factor; the preparation method of the white-cloud ginseng polysaccharide comprises the following steps:
1) Pulverizing radix Ginseng alba, and defatting with methanol at 60 deg.C;
2) Extracting with water at 80 deg.C for 3 hr for 2 times after degreasing, mixing extractive solutions, and concentrating under reduced pressure;
3) Precipitating the concentrated extractive solution with 80% ethanol, centrifuging, and removing supernatant;
4) Removing protein from the crude polysaccharide after alcohol precipitation by a sevege method, centrifuging, and discarding the precipitate for 5 times until no precipitate exists.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010078020.6A CN111297887B (en) | 2020-02-02 | 2020-02-02 | Preparation method and application of liver-protecting active component of Yunnan ginseng |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010078020.6A CN111297887B (en) | 2020-02-02 | 2020-02-02 | Preparation method and application of liver-protecting active component of Yunnan ginseng |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN111297887A CN111297887A (en) | 2020-06-19 |
| CN111297887B true CN111297887B (en) | 2023-04-18 |
Family
ID=71147017
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010078020.6A Active CN111297887B (en) | 2020-02-02 | 2020-02-02 | Preparation method and application of liver-protecting active component of Yunnan ginseng |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111297887B (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1648136A (en) * | 2005-01-12 | 2005-08-03 | 贵州省中国科学院天然产物化学重点实验室 | Companumoea root polyose, derivative and its preparing method and use |
| CN101966199A (en) * | 2010-09-27 | 2011-02-09 | 贵州省中国科学院天然产物化学重点实验室 | Application of campanumoea polysaccharide in preparation of medicament and health care food |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN118063635A (en) * | 2018-05-15 | 2024-05-24 | 云南云河药业股份有限公司 | Method for extracting active polysaccharide from white cloud ginseng |
-
2020
- 2020-02-02 CN CN202010078020.6A patent/CN111297887B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1648136A (en) * | 2005-01-12 | 2005-08-03 | 贵州省中国科学院天然产物化学重点实验室 | Companumoea root polyose, derivative and its preparing method and use |
| CN101966199A (en) * | 2010-09-27 | 2011-02-09 | 贵州省中国科学院天然产物化学重点实验室 | Application of campanumoea polysaccharide in preparation of medicament and health care food |
Also Published As
| Publication number | Publication date |
|---|---|
| CN111297887A (en) | 2020-06-19 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Chen et al. | Astragali Radix (Huangqi): A promising edible immunomodulatory herbal medicine | |
| Geng et al. | Preventive and therapeutic effect of Ganoderma lucidum on kidney injuries and diseases | |
| Gao et al. | Curcumae rhizoma and its major constituents against hepatobiliary disease: Pharmacotherapeutic properties and potential clinical applications | |
| CN111297885B (en) | Application of geranium polysaccharide in preparing liver-protecting medicine | |
| CN110772547B (en) | Application of Wenwangyibi extract in preparing medicine for treating hepatitis | |
| CN107951980B (en) | Application of hosta plantaginea flower and extract thereof in preparing medicine for treating chronic prostatitis | |
| CN117503782A (en) | Application of ginsenoside CK in preparing medicine for preventing and treating immune hepatitis | |
| CN111297887B (en) | Preparation method and application of liver-protecting active component of Yunnan ginseng | |
| WO2017121333A1 (en) | Use of cistanche tubulosa extract and isoacteoside in protection of muscles | |
| CN101433595A (en) | Desertliving cistanche alcohol extract as well as preparation method and use thereof | |
| CN115006494B (en) | Inula flower composite anti-alcohol composition with pulse-displaying function, anti-alcohol and liver-protecting preparation and application thereof | |
| WO2013115534A1 (en) | Composition for preventing or treating multiple sclerosis | |
| EP3639835B1 (en) | Pharmaceutical composition consisting of actein and deoyxactein as active compounds | |
| CN113402585A (en) | Oyster peptide and application thereof in alcoholic liver injury | |
| KR100685472B1 (en) | Composition using vinegar processed ginseng preparation for treatment and prevention of type ? diabetes and metabolic syndrome | |
| CN102940621B (en) | Application of methyl ferulic acid in preparation of medicine for preventing and curing hepatic fibrosis | |
| CN112515167A (en) | Pulse-developing inula flower composite extract and preparation method and application thereof | |
| CN111297886A (en) | Application of gentian polysaccharide in preparation of medicine for preventing or treating liver injury diseases | |
| CN106822152B (en) | Pharmaceutical composition and application thereof | |
| CN111346102A (en) | Application of baicalin in treating and preventing non-alcoholic fatty liver disease | |
| CN104510857B (en) | A kind of Chinese medicinal effective-part composition for blood fat reducing and preparation thereof | |
| CN113332416B (en) | Application of glutamine dipeptide in preparation of medicine for treating non-alcoholic fatty liver disease | |
| CN106729577B (en) | Traditional Chinese medicine composition for treating rheumatism and preparation method thereof | |
| CN107233378B (en) | A pharmaceutical composition for treating hepatitis and/or hepatic fibrosis, and its preparation method | |
| CN112089725A (en) | Application of aster polysaccharide in preparation of medicine for treating and/or preventing liver diseases |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |