Cardamine violifolia extract and application thereof
By using the extract of the sacred leaf crust extract, problems such as intestinal flora disorders, intestinal damage and blood function damage caused by drugs or chemical reagents are solved, and effective prevention and treatment of these symptoms are achieved.
Patent Information
- Application Number
- CN202411638609.1
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-11-15
- Publication Date
- 2025-02-11
AI Technical Summary
The prior art is difficult to effectively prevent and treat symptoms such as intestinal flora disorders, intestinal damage and blood function damage caused by drugs or chemical agents such as carbon tetrachloride.
The extract of the sacred leaf crust and shepherd's leaves is prepared by enzymatic method. It is used to prepare dry extract or dry extract for the preparation of compositions or preparations for the prevention and treatment of intestinal microbial disorders, hypoglycemia, hypoalbuminemia, Alanine transaminase and cerealis transaminase activity, hyperlipidemia and intestinal damage.
The extract of the sacred leaf crumbled rice shepherd's extract significantly improves intestinal flora disorders, increases blood sugar and albumin levels, reduces gluten-transferase activity, reduces hyperlipidemia and intestinal damage, and has excellent preventive and therapeutic effects.
Smart Images
Figure CN119405708A_ABST
Abstract
Description
Technical Field
[0001] The invention relates to the field of medicines, and in particular to a Cardamine violaceum extract and a use thereof. Background Art
[0002] Drugs or chemical reagents often have toxic side effects, and their application or poisoning often causes damage to the digestive system and blood system with vigorous metabolism. For example, carbon tetrachloride is a commonly used chemical reagent in the field of pharmaceutical chemical industry, and is often used as a solvent for drug organic synthesis or as an extractant for drug extraction. However, contact, inhalation, application or accidental ingestion of carbon tetrachloride chemical reagents often causes carbon tetrachloride poisoning, which often leads to adverse symptoms in the digestive system and blood system with vigorous metabolism, such as intestinal flora imbalance, intestinal damage and functional damage of the blood system such as decreased blood sugar and albumin levels, abnormal enzyme activity in the blood, etc., which in turn lead to discomfort in the body such as nausea, vomiting, indigestion, fatigue and edema, etc. In severe cases, it can further lead to the occurrence of acute symptoms such as acute enteritis and shock, and even endanger life and health. The effective prevention and treatment of symptoms such as intestinal flora imbalance, intestinal damage and blood function damage caused by carbon tetrachloride chemical reagent poisoning is of great significance to improving carbon tetrachloride chemical reagent poisoning. Therefore, how to prevent and treat damage caused by the toxic side effects of drugs or chemical reagents, such as intestinal flora imbalance, intestinal damage and blood function damage, has always been a research hotspot today.
[0003] Therefore, there is a need in the art to develop a drug that can effectively treat the damage caused by the toxic side effects of drugs or chemical agents, such as intestinal flora imbalance, intestinal damage, and blood function damage. Summary of the invention
[0004] The object of the present invention is to provide a use of a Cardamine violacea extract, wherein the Cardamine violacea extract can be used to improve intestinal flora imbalance, and has excellent preventive and therapeutic effects on hypoglycemia, hypoalbuminemia, elevated alanine aminotransferase and aspartate aminotransferase activities, hyperlipidemia and intestinal damage.
[0005] In a first aspect, the present invention provides a use of an extract of Cardamine violaceum for preparing a composition or preparation, wherein the composition or preparation is used for one or more uses selected from the following groups: (i) preventing and / or treating intestinal dysbiosis; (ii) preventing and / or treating hypoglycemia; (iii) increasing blood sugar; (iv) preventing and / or treating hypoalbuminemia; (v) increasing albumin; (vi) preventing and / or treating elevated alanine aminotransferase activity; (vii) reducing alanine aminotransferase activity; (viii) preventing and / or treating elevated aspartate aminotransferase activity; (ix) reducing aspartate aminotransferase activity; (x) preventing and / or treating hyperlipidemia; (xi) reducing triglycerides; (xii) preventing and / or treating intestinal damage; and / or (xiii) preventing and / or treating carbon tetrachloride poisoning or its complications.
[0006] Preferably, the Cardamine violacea extract is a dry extract.
[0007] Preferably, the Cardamine violacea extract is a dry extract.
[0008] Preferably, the Cardamine violacea extract is prepared by the following method:
[0009] Cardamine violacea is enzymatically hydrolyzed with an aqueous solution containing an enzyme to obtain a Cardamine violacea extract.
[0010] Preferably, the enzyme comprises one or more of cellulase, hemicellulase and xylanase.
[0011] Preferably, the enzymes include cellulases, hemicellulases and xylanases.
[0012] Preferably, the amount of the cellulase is 0.5-8% by weight of Cardamine violacea, preferably 1-6%, more preferably 1-5%, more preferably 2-4%, more preferably 2.5-3.5%, more preferably 2.8-3.2%, and most preferably 3.0%.
[0013] Preferably, the amount of the hemicellulase is 0.2-5.0% by weight of Cardamine violacea, preferably 0.2-3.0%, more preferably 0.3-2.0%, more preferably 0.5-1.5%, more preferably 0.8-1.2%, and most preferably 1.0%.
[0014] Preferably, the amount of xylanase used is 0.02-1.0% of the weight of Cardamine violacea, preferably 0.02-0.5%, more preferably 0.02-0.3%, more preferably 0.03-0.2%, more preferably 0.05-0.15%, more preferably 0.08-0.12%, and most preferably 0.1%.
[0015] Preferably, the pH of the enzyme-containing aqueous solution is 3.0-7.0, more preferably 3.0-6.5, more preferably 4-6, more preferably 4.5-5.5, more preferably 4.8-5.2, and most preferably 5.0.
[0016] Preferably, the pH of the aqueous solution containing the enzyme is adjusted with acetic acid.
[0017] Preferably, the pH of the aqueous solution containing the enzyme is adjusted to 3.0-7.0 with acetic acid, preferably 3.0-6.5, more preferably 4-6, more preferably 4.5-5.5, more preferably 4.8-5.2, and most preferably 5.0.
[0018] Preferably, the weight ratio of the enzyme-containing aqueous solution to the Cardamine violaceae is (10-45):1, preferably (10-40:1, more preferably (15-35:1), more preferably (20-30:1, more preferably (23-27):1, and most preferably 25:1.
[0019] Preferably, the temperature of the enzymatic hydrolysis is 35-55°C, more preferably 40-55°C, more preferably 45-55°C, more preferably 48-52°C, and most preferably 50°C.
[0020] Preferably, the enzymatic hydrolysis time is 1-4 h, more preferably 1-3 h, more preferably 1.5-2.5 h, more preferably 1.8-2.2 h, and most preferably 2.0 h.
[0021] Preferably, the enzymatic hydrolyzate after enzymatic hydrolysis is filtered.
[0022] Preferably, the enzymatic hydrolyzate after enzymatic hydrolysis is filtered to obtain a filtrate, and the Cardamine violaceum extract is separated from the filtrate.
[0023] Preferably, the filtration comprises filtering with a microfiltration membrane and an ultrafiltration membrane in sequence.
[0024] Preferably, the pore size of the microfiltration membrane is 1 μm.
[0025] Preferably, the pore size of the ultrafiltration membrane is 1 nm.
[0026] Preferably, the separation comprises subjecting the filtrate to rotary vacuum evaporation to obtain the Cardamine violaceum extract.
[0027] Preferably, the Cardamine violacea extract is prepared by the following method:
[0028] Add 25 times the amount of water to Cardamine violacea, adjust the pH to 5.0 with acetic acid, add cellulase, hemicellulase and xylanase, stir and enzymolyze at 50°C for 2h, filter, and filter the obtained filtrate with a microfiltration membrane and an ultrafiltration membrane in turn, and obtain the filtrate after rotary vacuum evaporation to obtain the Cardamine violacea extract;
[0029] The dosage of cellulase, hemicellulase and xylanase is 3%, 1% and 0.1% of the weight of Cardamine violacea respectively;
[0030] The pore size of the microfiltration membrane is 1 μm, and the pore size of the ultrafiltration membrane is 1 nm.
[0031] Preferably, the composition or preparation is administered to a human or non-human mammal.
[0032] Preferably, the composition or preparation is administered to a subject to whom carbon tetrachloride is administered or poisoned by carbon tetrachloride.
[0033] Preferably, the subject of the prevention and / or treatment is a human or non-human mammal.
[0034] Preferably, the subject of the prevention and / or treatment is a subject administered with carbon tetrachloride or poisoned by carbon tetrachloride.
[0035] Preferably, the intestinal microecology is the intestinal microecology of a human or non-human mammal.
[0036] Preferably, the intestinal dysbiosis includes intestinal dysbiosis caused by administration of carbon tetrachloride or carbon tetrachloride poisoning.
[0037] Preferably, the intestinal microecological imbalance includes intestinal flora imbalance.
[0038] Preferably, the intestinal flora imbalance includes intestinal flora quantity imbalance.
[0039] Preferably, the intestinal flora includes one or more of Alloprevotella, Alistipes, Allobaculum, Eubacterium coprostanoligenes group, Rikenellaceae RC9 gut group, Harryflintia, Bacteroides, Bifidobacterium and Faecalibaculum.
[0040] Preferably, the intestinal dysbiosis includes intestinal hyperbacteriosis.
[0041] Preferably, the intestinal dysbiosis includes intestinal hyperbacteria, and the intestinal flora is selected from the following group: Alloprevotella, Alistipes, Allobaculum, Eubacterium_coprostanoligenes_group, Rikenellaceae_RC9_gut_group, Harryflintia, or a combination thereof.
[0042] Preferably, the intestinal dysbiosis comprises an intestinal flora hyperbiosis selected from the group consisting of Alloprevotella, Alistipes, Allobaculum, Eubacterium coprostanoligenes group, Rikenellaceae RC9 gut group, Harryflintia, or a combination thereof.
[0043] Preferably, the intestinal flora hyperbiosis includes intestinal flora overabundance disorder.
[0044] Preferably, the intestinal microecological imbalance includes intestinal flora deficiency.
[0045] Preferably, the intestinal dysbiosis includes intestinal flora deficiency, and the intestinal flora is selected from the following group: Bacteroides, Bifidobacterium, Faecalibaculum, or a combination thereof.
[0046] Preferably, the intestinal dysbiosis comprises intestinal flora deficiency selected from the group consisting of Bacteroides, Bifidobacterium, Faecalibaculum, or a combination thereof.
[0047] Preferably, the intestinal flora deficiency includes intestinal flora quantity deficiency.
[0048] Preferably, the prevention and / or treatment of intestinal dysbiosis comprises preventing and / or treating intestinal dysbiosis by reducing the number of intestinal flora selected from the group consisting of Alloprevotella, Alistipes, Allobaculum, Eubacterium_coprostanoligenes_group, Rikenellaceae_RC9_gut_group, Harryflintia, or a combination thereof.
[0049] Preferably, the prevention and / or treatment of intestinal dysbiosis comprises preventing and / or treating intestinal dysbiosis by increasing the number of intestinal flora selected from the group consisting of Bacteroides, Bifidobacterium, Faecalibaculum, or a combination thereof.
[0050] Preferably, the blood glucose is the blood glucose of a human or non-human mammal.
[0051] Preferably, the hypoglycemia is hypoglycemia in a human or non-human mammal.
[0052] Preferably, the hypoglycemia comprises hypoglycemia caused by administration of carbon tetrachloride or carbon tetrachloride poisoning.
[0053] Preferably, the prevention and / or treatment of hypoglycemia comprises preventing and / or treating hypoglycemia by increasing blood glucose.
[0054] Preferably, said increasing blood sugar comprises increasing the blood sugar content.
[0055] Preferably, the hypoalbuminemia is hypoalbuminemia in humans or non-human mammals.
[0056] Preferably, the hypoalbuminemia includes hypoalbuminemia caused by administration of carbon tetrachloride or carbon tetrachloride poisoning.
[0057] Preferably, the albumin is human or non-human mammal albumin.
[0058] Preferably, the albumin comprises albumin in blood.
[0059] Preferably, the prevention and / or treatment of hypoalbuminemia comprises preventing and / or treating hypoalbuminemia by increasing albumin.
[0060] Preferably, said increasing albumin comprises increasing the albumin content.
[0061] Preferably, the alanine aminotransferase is alanine aminotransferase from a human or non-human mammal.
[0062] Preferably, the increased activity of alanine aminotransferase includes increased activity of alanine aminotransferase caused by administration of carbon tetrachloride or carbon tetrachloride poisoning.
[0063] Preferably, the alanine aminotransferase comprises alanine aminotransferase in blood.
[0064] Preferably, the prevention and / or treatment of elevated alanine aminotransferase activity comprises preventing and / or treating elevated alanine aminotransferase activity by reducing the activity of alanine aminotransferase.
[0065] Preferably, the aspartate aminotransferase is aspartate aminotransferase from a human or non-human mammal.
[0066] Preferably, the increased aspartate aminotransferase activity includes the increased aspartate aminotransferase activity caused by the administration of carbon tetrachloride or carbon tetrachloride poisoning.
[0067] Preferably, the aspartate aminotransferase comprises aspartate aminotransferase in blood.
[0068] Preferably, the prevention and / or treatment of elevated aspartate aminotransferase activity comprises preventing and / or treating elevated aspartate aminotransferase activity by reducing aspartate aminotransferase activity.
[0069] Preferably, the hyperlipidemia is hyperlipidemia in humans or non-human mammals.
[0070] Preferably, the hyperlipidemia includes high triglycerides.
[0071] Preferably, the hyperlipidemia includes hyperlipidemia caused by administration of carbon tetrachloride or carbon tetrachloride poisoning.
[0072] Preferably, the triglyceride is a triglyceride from a human or non-human mammal.
[0073] Preferably, the triglycerides include triglycerides in blood.
[0074] Preferably, said reducing triglycerides comprises reducing the content of triglycerides.
[0075] Preferably, the prevention and / or treatment of hyperlipidemia comprises preventing and / or treating hyperlipidemia by lowering triglycerides.
[0076] Preferably, the subject of the intestinal injury is intestinal injury of a human or non-human mammal.
[0077] Preferably, the intestinal damage includes intestinal damage caused by administration of carbon tetrachloride or carbon tetrachloride poisoning.
[0078] Preferably, the intestinal damage includes one or more characteristics selected from the following group: intestinal mucosal disorder, intestinal mucosal thickening, increased intestinal inflammatory cells, severe intestinal extracellular deposition, or a combination thereof.
[0079] Preferably, the intestine comprises the colon.
[0080] Preferably, the prevention and / or treatment of intestinal injury comprises preventing and / or treating intestinal injury by improving one or more characteristics selected from the following group: intestinal mucosal disorder, intestinal mucosal thickening, increased intestinal inflammatory cells, severe intestinal extracellular deposition, or a combination thereof
[0081] Preferably, the subject of carbon tetrachloride poisoning or its complications is carbon tetrachloride poisoning or its complications in humans or non-human mammals.
[0082] Preferably, the complications of carbon tetrachloride poisoning are selected from the following group: intestinal dysbiosis, hypoglycemia, hypoalbuminemia, elevated alanine aminotransferase activity, elevated aspartate aminotransferase activity, hyperlipidemia, intestinal damage, or a combination thereof.
[0083] Preferably, the prevention and / or treatment of carbon tetrachloride poisoning or its complications includes (i) prevention and / or treatment of intestinal microecological imbalance; (ii) prevention and / or treatment of hypoglycemia; (iii) increase of blood glucose; (iv) prevention and / or treatment of hypoalbuminemia; (v) increase of albumin; (vi) prevention and / or treatment of increased alanine aminotransferase activity; (vii) reduction of alanine aminotransferase activity; (viii) prevention and / or treatment of increased aspartate aminotransferase activity; (ix) reduction of aspartate aminotransferase activity; (x) prevention and / or treatment of hyperlipidemia; (xi) reduction of triglycerides; and / or (xii) prevention and / or treatment of intestinal damage.
[0084] Preferably, the subject comprises a subject to which carbon tetrachloride is administered.
[0085] Preferably, the subject comprises a subject poisoned by carbon tetrachloride.
[0086] Preferably, the subject includes a subject administered with carbon tetrachloride or poisoned by carbon tetrachloride.
[0087] Preferably, the subject comprises a subject suffering from carbon tetrachloride poisoning or its complications.
[0088] Preferably, the subject comprises a subject with intestinal damage.
[0089] Preferably, the intestinal damage includes intestinal damage caused by administration of carbon tetrachloride or carbon tetrachloride poisoning.
[0090] Preferably, the carbon tetrachloride poisoning includes carbon tetrachloride poisoning caused by oral ingestion and / or inhalation of carbon tetrachloride.
[0091] Preferably, the subject includes a subject suffering from intestinal dysbiosis, hypoglycemia, hypoalbuminemia, elevated alanine aminotransferase activity, elevated aspartate aminotransferase activity, hyperlipidemia, intestinal damage, and / or carbon tetrachloride poisoning or its complications.
[0092] Preferably, the non-human mammal is a mouse, rat, rabbit, cow, sheep, dog or horse.
[0093] Preferably, the mice comprise C57BL / 6 mice.
[0094] Preferably, the composition or preparation is a pharmaceutical composition or pharmaceutical preparation.
[0095] Preferably, the composition or preparation further comprises a pharmaceutically acceptable carrier.
[0096] Preferably, the composition or preparation is in the form of a solid preparation, a semisolid preparation or a liquid preparation.
[0097] Preferably, the composition or preparation is in the form of an oral preparation, an external preparation or an injection preparation.
[0098] Preferably, the composition or preparation is in the form of emulsion, tablet, capsule, oral liquid, granule, powder, syrup or injection.
[0099] In a second aspect, the present invention provides a method for (i) preventing and / or treating intestinal dysbiosis; (ii) preventing and / or treating hypoglycemia; (iii) increasing blood sugar; (iv) preventing and / or treating hypoalbuminemia; (v) increasing albumin; (vi) preventing and / or treating elevated alanine aminotransferase activity; (vii) reducing alanine aminotransferase activity; (viii) preventing and / or treating elevated aspartate aminotransferase activity; (ix) reducing aspartate aminotransferase activity; (x) preventing and / or treating hyperlipidemia; (xi) reducing triglycerides; (xii) preventing and / or treating intestinal damage; and / or (xiii) preventing and / or treating carbon tetrachloride poisoning or its complications, the method comprising administering an extract of Cardamine violaceum to a subject in need thereof.
[0100] Preferably, the subject is a human or non-human mammal.
[0101] Preferably, the non-human mammal is a mouse, rat, rabbit, cow, sheep, dog or horse.
[0102] Preferably, the mice comprise C57BL / 6 mice.
[0103] Preferably, the administration is oral administration, external administration or injection administration.
[0104] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (such as embodiments) can be combined with each other to form a new or preferred technical solution. Due to space limitations, they will not be described one by one here. BRIEF DESCRIPTION OF THE DRAWINGS
[0105] Figure 1 Figure 2 is the hematoxylin-eosin (HE) staining of mouse colon tissues in the normal control group, vehicle group, model group and Cardamine violaceum extract group, where a is the normal control group, b is the vehicle group, c is the model group, and f is the Cardamine violaceum extract group.
[0106] Figure 2 The t-test was used to detect the differences in intestinal microorganisms between the vehicle group and the model group.
[0107] Figure 3 The t-test was used to detect the differences in intestinal microorganisms between the model group and the Cardamine violaceum extract group. DETAILED DESCRIPTION
[0108] The present invention unexpectedly discovered that the Cardamine violacea extract can be used to improve intestinal flora imbalance, has excellent preventive and therapeutic effects on hypoglycemia, hypoalbuminemia, elevated alanine aminotransferase and aspartate aminotransferase activities, hyperlipidemia and intestinal damage, and can be used to prevent and treat damage caused by the toxic side effects of drugs or chemical reagents such as carbon tetrachloride, such as intestinal flora imbalance, intestinal damage and blood function damage.
[0109] the term
[0110] As used herein, the terms "including", "comprising", and "containing" are used interchangeably and include not only open definitions, but also semi-closed and closed definitions, and the terms include "consisting of", "consisting essentially of".
[0111] In the present invention, the Latin name of the term "Cardamine violifolia" is Cardamine violifolia.
[0112] In the present invention, the term "PBS" refers to phosphate buffered saline.
[0113] In the present invention, the term "carbon tetrachloride" is Tetrachloromethane, abbreviated as CCl4.
[0114] In the present invention, the term "albumin" is referred to as albumin in English.
[0115] In the present invention, the term "alanine aminotransferase" is Alanine aminotransferas in English.
[0116] In the present invention, the term "aspartate aminotransferase" is Aspartate aminotransferase in English.
[0117] In the present invention, the term "triglyceride" in English is triglyceride.
[0118] In the present invention, the term "prevention" refers to a method of preventing the onset of a disease and / or its attendant symptoms or protecting a subject from acquiring a disease.
[0119] In the present invention, the term "treatment" includes delaying and stopping the progression of the disease, or eliminating the disease, and does not require 100% inhibition, elimination and reversal.
[0120] Cardamine Viola Extract
[0121] The present invention provides a Cardamine violaceae extract, which is prepared by the following method:
[0122] Cardamine violacea is enzymatically hydrolyzed with an aqueous solution containing an enzyme to obtain a Cardamine violacea extract.
[0123] Specifically, the Cardamine viola extract of the present invention is as described above in the first aspect of the present invention.
[0124] use
[0125] The present invention provides a use of an extract of Cardamine violaceum for preparing a composition or a preparation, wherein the composition or the preparation is used for one or more uses selected from the following groups: (i) preventing and / or treating intestinal microecological imbalance; (ii) preventing and / or treating hypoglycemia; (iii) increasing blood sugar; (iv) preventing and / or treating hypoalbuminemia; (v) increasing albumin; (vi) preventing and / or treating elevated alanine aminotransferase activity; (vii) reducing alanine aminotransferase activity; (viii) preventing and / or treating elevated aspartate aminotransferase activity; (ix) reducing aspartate aminotransferase activity; (x) preventing and / or treating hyperlipidemia; (xi) reducing triglycerides; (xii) preventing and / or treating intestinal damage; and / or (xiii) preventing and / or treating carbon tetrachloride poisoning or its complications.
[0126] Specifically, the use of the Cardamine viola extract of the present invention is as described above in the first aspect of the present invention.
[0127] Compositions, formulations and kits
[0128] The present invention also provides a composition or preparation, which is preferably a pharmaceutical composition or preparation. The composition or preparation of the present invention further comprises a pharmaceutically acceptable carrier.
[0129] As used herein, the term "pharmaceutically acceptable carrier" refers to one or more compatible solid, semisolid, liquid or gel fillers that are suitable for human or animal use and have sufficient purity and sufficiently low toxicity.
[0130] It should be understood that in the present invention, the carrier is not particularly limited, and can be selected from materials commonly used in the art, or prepared by conventional methods, or purchased from the market. Examples of pharmaceutically acceptable carriers include cellulose and its derivatives (such as methylcellulose, ethylcellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose, etc.), gelatin, talcum powder, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, etc.), polyols (such as propylene glycol, glycerol, sorbitol, etc.), emulsifiers (such as Tween), wetting agents (such as sodium lauryl sulfate), buffers, chelating agents, thickeners, pH regulators, transdermal enhancers, colorants, flavoring agents, stabilizers, antioxidants, preservatives, antibacterial agents, pyrogen-free water, etc.
[0131] In the present invention, the dosage form of the composition or preparation includes (but is not limited to) solid preparation, semisolid preparation or liquid preparation.
[0132] In the present invention, the dosage form of the composition or preparation includes (but is not limited to) oral preparations, injection preparations or external preparations.
[0133] Typically, the injection preparation is an intravenous injection preparation, an intramuscular injection preparation, a subcutaneous injection preparation, an intradermal injection preparation or an intraperitoneal injection preparation.
[0134] The pharmaceutical preparation should be matched with the mode of administration. When administering, a therapeutically effective amount of the drug is administered to the desired subject (such as a human or non-human mammal). As used herein, the term "therapeutically effective amount" refers to an amount that produces a function or activity on humans and / or animals and can be accepted by humans and / or animals. It should be understood by those of ordinary skill in the art that the "therapeutically effective amount" may vary depending on the form of the pharmaceutical preparation, the route of administration, the adjuvants of the drug used, the severity of the disease, and the combination of other drugs, etc., which are all within the skill of skilled physicians.
[0135] The main technical effects achieved by the present invention include:
[0136] The present invention finds that the Cardamine violacea extract can be used to improve intestinal flora imbalance, has excellent prevention and treatment effects on hypoglycemia, hypoalbuminemia, elevated alanine aminotransferase and aspartate aminotransferase activities, hyperlipidemia and intestinal damage, and can be used to prevent and treat damage caused by the toxic side effects of drugs or chemical reagents such as carbon tetrachloride, such as intestinal flora imbalance, intestinal damage and blood function damage.
[0137] The present invention is further described below in conjunction with specific examples. It should be understood that the following specific examples are based on the technical solution and provide detailed implementation methods and specific operation processes, but the protection scope of the present invention is not limited to these examples.
[0138] Example 1
[0139] 1. Preparation of Cardamine Viola Extract:
[0140] Add 25 times the amount of water to the crushed Cardamine violacea, adjust the pH to 5.0 with acetic acid, add cellulase, hemicellulase and xylanase (wherein, the amount of cellulase, hemicellulase and xylanase is 3%, 1% and 0.1% of the weight of Cardamine violacea, respectively), stir and enzymolyze at 50°C for 2h, filter, and filter the obtained filtrate with a microfiltration membrane (membrane pore size of 1μm) and an ultrafiltration membrane (membrane pore size of 1nm) in turn, and obtain the filtrate. After rotary reduced pressure evaporation, the Cardamine violacea extract is obtained.
[0141] 2. Preparation of the drug solution:
[0142] 20% carbon tetrachloride oil solution: Mix 1 ml of carbon tetrachloride and 4 ml of olive oil to obtain a 20% carbon tetrachloride oil solution.
[0143] Cardamine violaceae extract dispersion: the Cardamine violaceae extract was dispersed in PBS 7.4 buffer to obtain the Cardamine violaceae extract dispersion.
[0144] 3. Experimental Methods
[0145] Normal healthy male C57BL / 6 mice were divided into normal control group, vehicle group, model group and Cardamine violata extract group, with 6 mice in each group, and the administration method was as follows:
[0146] Vehicle group: Normal saline was given by gavage once a day for 56 days from the beginning of the experiment. Olive oil was injected intraperitoneally starting from the 15th day (dosage volume was 0.004 ml / g). Olive oil was given 3 times a week for 4 consecutive weeks. The experiment was terminated on the 57th day.
[0147] Model group: Normal saline was gavaged once a day for 56 days from the beginning of the experiment. Starting from the 15th day, 20% carbon tetrachloride oil solution was injected intraperitoneally (the dosage was 0.005 ml / g, calculated as 20% carbon tetrachloride oil solution). The 20% carbon tetrachloride oil solution was administered 3 times / week for 4 consecutive weeks. The experiment was terminated on the 57th day.
[0148] Cardamine violeta extract group: Cardamine violeta extract dispersion was administered intragastrically (dosage was 0.8 mg / kg, calculated as Cardamine violeta extract) once a day within 14 days from the beginning of the experiment. Starting from the 15th day, 20% carbon tetrachloride oil solution was injected intraperitoneally (dosage was 0.005 ml / g, calculated as 20% carbon tetrachloride oil solution), and at the same time, a dispersion of Cardamine violaceae extract was administered by gavage (dosage was 0.8 mg / kg, calculated as Cardamine violaceae extract). The 20% carbon tetrachloride oil solution and the dispersion of Cardamine violaceae extract were administered 3 times / week, respectively, and the administration was continued for 4 weeks. On the 43rd day, the administration of 20% carbon tetrachloride oil solution was stopped, and the dispersion of Cardamine violaceae extract (dosage was 0.8 mg / kg, calculated as Cardamine violaceae extract) was continued by gavage for 2 weeks (3 times / week), and the experiment was terminated on the 57th day.
[0149] Normal control group: The administration method of the normal control group was the same as that of the Cardamine violaceae extract group, except that the Cardamine violaceae extract dispersion and 20% carbon tetrachloride oil solution were replaced by the same volume of physiological saline, and the experiment was terminated on the 57th day.
[0150] 4. Experimental Results
[0151] 4.1 At the end of the 57th day, the colons of mice in the normal control group, vehicle group, model group and Cardamine violaceum extract group were fixed in a centrifuge tube filled with 4% paraformaldehyde solution. After paraformaldehyde fixation, they were first embedded in paraffin and then dehydrated and sectioned for hematoxylin-eosin (HE) staining. The results are shown in Figure 4. Figure 1 shown.
[0152] from Figure 1 It can be seen that compared with the vehicle group given olive oil, the mice in the model group given carbon tetrachloride had disordered and thickened colon mucosa, increased inflammatory cells, and severe extracellular deposition, indicating that carbon tetrachloride caused intestinal damage. Compared with the carbon tetrachloride model group, the disorder and thickening of the colon mucosa in the mice in the group given Cardamine violaceum extract were effectively alleviated, and the inflammatory cells and extracellular deposition were significantly reduced, indicating that the Cardamine violaceum extract can effectively improve and treat intestinal damage caused by carbon tetrachloride.
[0153] 4.2 At the end of the 57th day, blood was collected from the eyeballs of mice in the normal control group, solvent group, model group and Cardamine violaceum extract group, placed in labeled and anticoagulant-rinsed 1.5 ml centrifuge tubes, placed in a 4°C refrigerator for 12 hours, and centrifuged in a high-speed desktop refrigerated centrifuge at 4°C and 3500g / min for 10 minutes. After centrifugation, the supernatant was aspirated with a pipette on an ice box and transferred to a pre-cooled clean centrifuge tube. The fasting glucose (Glu-G), triglyceride (TG) and albumin (ALB) levels in serum and the activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were detected using an automatic biochemical analyzer. The results are shown in Table 1 below.
[0154] Table 1 Detection of serum indicators in each group
[0155]
[0156] Note: Compared with the vehicle group, *p<0.05, **p<0.01; compared with the model group, # p<0.05, ## p<0.01.
[0157] As can be seen from Table 1, compared with the vehicle group given olive oil, the fasting glucose (Glu-G) content in the serum of the model group mice given carbon tetrachloride was reduced, the activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were increased, the content of triglyceride (TG) was increased, and the content of albumin (ALB) was reduced. Therefore, carbon tetrachloride can cause a decrease in blood glucose (Glu-G) content, an increase in the activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST), an increase in the content of triglyceride (TG), and a decrease in the content of albumin (ALB), thereby leading to hypoglycemia, alanine aminotransferase (ALT) and aspartate aminotransferase (AST), and a decrease in the content of triglyceride (TG). However, compared with the model group given carbon tetrachloride, the Cardamine violeta extract was able to increase the serum glucose (Glu-G) content, reduce the serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities, reduce the triglyceride (TG) content and increase the albumin (ALB) content after administration, indicating that the Cardamine violeta extract has an excellent therapeutic effect on hypoglycemia, elevated alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities, hyperlipidemia and hypoalbuminemia.
[0158] 4.3 At the end of the 57th day, the colon contents of the mice in the vehicle group, model group and Cardamine violaceum extract group (10 samples in each group) were collected in sterile cryopreservation tubes, and DNA was extracted from the frozen contents and 16S rDNA was sequenced to determine the changes in the intestinal flora of the mice. The results are as follows: Figure 2 and Figure 3 shown.
[0159] from Figure 2 It can be seen that compared with the vehicle group, the abundance of Alistipes and Rikenellaceae_RC9_gut_group in the fecal flora of the model group given carbon tetrachloride was significantly increased. Figure 3 It can be seen that compared with the model group given carbon tetrachloride, the fecal flora of the mice in the Cardamine violet extract group treated with Cardamine violet extract was higher in Alloprevotella, Alistipes, Allobaculum, Eubacterium_coprostanoligenes_group, Rikenellaceae_RC9_gut_group, and Rikenellaceae_RC9_gut_group. The abundance of Bacteroides, Bifidobacterium and Faecalibaculum was significantly increased, indicating that the extract of Cardamine violaceum can effectively reduce the abundance of Alloprevotella, Alistipes, Allobaculum, Eubacterium_coprostano in the intestine. ligenes_group (coprostanol-producing true bacteria), Rikenellaceae_RC9_gut_group (Riken bacteria genus_rc9_intestinal family) and Harryflintia abundance and increase the abundance of Bacteroides (Bacteroides), Bifidobacterium (Bifidobacterium) and Faecalibaculum (Faecalibaculum) in the intestine, for the prevention and treatment of intestinal Alloprevotella (Pseudo-Prevotella), Alistipes (Another branch of bacteria), Allobaculum (Another stick bacteria), Eubacterium_coprostanoligenes_group (coprostanol-producing true bacteria), Rikenellaceae_RC9_gut_group (Riken bacteria genus_rc9_intestinal family) and Harryflintia excess and Bacteroides (Bacteroides), Bifidobacterium (Bifidobacterium) and Faecalibaculum (Faecalibaculum) deficiency.
[0160] The above description is only the preferred embodiment of the present application and is not intended to limit the present application. For those skilled in the art, the present application may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application shall be included in the protection scope of the present application.
Claims
1. A use of Cardamine violacea extract, characterized in that: For preparing a composition or preparation, the composition or preparation is used for one or more purposes selected from the following group: (i) prevention and / or treatment of intestinal microecological imbalance; (ii) prevention and / or treatment of hypoglycemia; (iii) increase of blood sugar; (iv) prevention and / or treatment of hypoalbuminemia; (v) increase of albumin; (vi) prevention and / or treatment of elevated alanine aminotransferase activity; (vii) reduction of alanine aminotransferase activity; (viii) prevention and / or treatment of elevated aspartate aminotransferase activity; (ix) reduction of aspartate aminotransferase activity; (x) prevention and / or treatment of hyperlipidemia; (xi) reduction of triglycerides; (xii) prevention and / or treatment of intestinal damage; and / or (xiii) prevention and / or treatment of carbon tetrachloride poisoning or its complications; The Cardamine violacea extract is prepared by the following method: Cardamine violacea is enzymatically hydrolyzed with an aqueous solution containing an enzyme to obtain a Cardamine violacea extract.
2. The use according to claim 1, characterized in that The Cardamine violacea extract is prepared by the following method: Add 25 times the amount of water to Cardamine violacea, adjust the pH to 5.0 with acetic acid, add cellulase, hemicellulase and xylanase, stir and enzymolyze at 50°C for 2h, filter, and filter the obtained filtrate with a microfiltration membrane and an ultrafiltration membrane in turn, and obtain the filtrate after rotary vacuum evaporation to obtain the Cardamine violacea extract; The dosage of cellulase, hemicellulase and xylanase is 3%, 1% and 0.1% of the weight of Cardamine violacea respectively; The pore size of the microfiltration membrane is 1 μm, and the pore size of the ultrafiltration membrane is 1 nm.
3. The use according to claim 1, characterized in that The intestinal flora imbalance includes intestinal flora quantity imbalance, and the intestinal flora includes one or more of Alloprevotella, Alistipes, Allobaculum, Eubacterium_coprostanoligenes_group, Rikenellaceae_RC9_gut_group, Harryflintia, Bacteroides, Bifidobacterium and Faecalibaculum.
4. The use according to claim 1, characterized in that The intestinal dysbiosis includes intestinal flora hyperbiosis selected from the following groups: Alloprevotella, Alistipes, Allobaculum, Eubacterium_coprostanoligenes_group, Rikenellaceae_RC9_gut_group, Harryflintia, or a combination thereof.
5. The use according to claim 1, characterized in that The intestinal microecological imbalance includes intestinal flora deficiency selected from the group consisting of Bacteroides, Bifidobacterium, Faecalibaculum, or a combination thereof.
6. The use according to claim 1, characterized in that The intestinal microecological imbalance includes intestinal microecological imbalance caused by administration of carbon tetrachloride or carbon tetrachloride poisoning; The hypoglycemia includes hypoglycemia caused by administration of carbon tetrachloride or carbon tetrachloride poisoning; The hypoalbuminemia includes hypoalbuminemia caused by administration of carbon tetrachloride or carbon tetrachloride poisoning; The increased activity of alanine aminotransferase includes the increased activity of alanine aminotransferase caused by administration of carbon tetrachloride or carbon tetrachloride poisoning; The increased activity of aspartate aminotransferase includes the increased activity of aspartate aminotransferase caused by the administration of carbon tetrachloride or carbon tetrachloride poisoning; The hyperlipidemia includes hyperlipidemia caused by administration of carbon tetrachloride or carbon tetrachloride poisoning; and / or The intestinal damage includes intestinal damage caused by administration of carbon tetrachloride or carbon tetrachloride poisoning.
7. The use according to claim 1, characterized in that The complications of carbon tetrachloride poisoning are selected from the following group: intestinal dysbiosis, hypoglycemia, hypoalbuminemia, elevated alanine aminotransferase activity, elevated aspartate aminotransferase activity, hyperlipidemia, intestinal damage, or a combination thereof.
8. The use according to claim 1, characterized in that The composition or preparation is a pharmaceutical composition or pharmaceutical preparation.
9. The use according to claim 1, characterized in that The dosage form of the composition or preparation is an oral preparation, an external preparation or an injection preparation.
10. A method for (i) preventing and / or treating intestinal dysbiosis; (ii) preventing and / or treating hypoglycemia; (iii) increasing blood glucose; (iv) preventing and / or treating hypoalbuminemia; (v) increasing albumin; (vi) preventing and / or treating elevated alanine aminotransferase activity; (vii) reducing alanine aminotransferase activity; (viii) preventing and / or treating elevated aspartate aminotransferase activity; (ix) reducing aspartate aminotransferase activity; (x) preventing and / or treating hyperlipidemia; (xi) reducing triglycerides; (xii) preventing and / or treating intestinal damage; and / or (xiii) preventing and / or treating carbon tetrachloride poisoning or its complications, the method comprising administering an extract of Cardamine violaceum to a subject in need thereof.