CN113134020A - Preparation method and application of Mongolian medicine tribulus terrestris ethanol extract - Google Patents
Preparation method and application of Mongolian medicine tribulus terrestris ethanol extract Download PDFInfo
- Publication number
- CN113134020A CN113134020A CN202110529731.5A CN202110529731A CN113134020A CN 113134020 A CN113134020 A CN 113134020A CN 202110529731 A CN202110529731 A CN 202110529731A CN 113134020 A CN113134020 A CN 113134020A
- Authority
- CN
- China
- Prior art keywords
- ethanol extract
- tribulus terrestris
- model
- ethanol
- preparation
- 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.)
- Pending
Links
Images
Classifications
-
- 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)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
- A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
- A61P39/06—Free radical scavengers or antioxidants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P7/00—Drugs for disorders of the blood or the extracellular fluid
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization
Abstract
The invention discloses a preparation method and application of a Mongolian medicine tribulus terrestris ethanol extract, belonging to the field of medicines. The invention verifies that the caltrop ethanol extract can reduce hyperglycemia induced by high-fat feed in a manner of intragastric administration through a high-fat feed induced rat hyperglycemia model. Experimental results show that for hyperglycemia induced by high-fat feed, the tribulus terrestris ethanol extract can improve metabolic component indexes to be close to normal values, improve indexes of antioxidase SOD, CAT and GSH-Px, reduce serum MDA indexes and reduce the apoptosis rate of liver cells. The results show that the tribulus terrestris ethanol extract can reduce blood sugar through oxidation resistance and hepatocyte apoptosis expression reduction. The invention obtains a group of 18 biomarkers related to the caltrop ethanol extract by screening with a metabonomic method, and can play a role in the subsequent separation, research and identification of effective components in the caltrop ethanol extract.
Description
Technical Field
The invention relates to the field of medicines, in particular to a preparation method and application of a Mongolian medicine tribulus terrestris ethanol extract.
Background
Diabetes is a group of metabolic diseases characterized by hyperglycemia. Hyperglycemia is caused by a defect in insulin secretion or an impaired biological action, or both. The chronic hyperglycemia results in chronic damage and dysfunction of various tissues, particularly eyes, kidneys, heart, blood vessels and nerves. The clinical manifestations are polydipsia, diuresis, polyphagia, fatigue, weakness, obesity, etc. Reducing or controlling blood sugar is an important measure for controlling the development of diabetes, and the most direct way in clinic at present is to reduce blood sugar by insulin injection, but the price is high and certain pain is brought to patients. Therefore, the search for oral hypoglycemic drugs with low cost and good effect is very necessary to prevent and treat diabetes.
Tribulus terrestris L is fruit of Tribulus terrestris of Tribulus of Zygophyllaceae, which is named as "Octopus inferior", and has different names of "color code" and "color code" with wide distribution, easy acquisition and low cost. The caltrop contains caltrop saponin, flavonoid, polysaccharide, alkaloid and rich trace elements, and no report related to the preparation of hypoglycemic drugs by using caltrop ethanol extracts exists at present.
Disclosure of Invention
The invention aims to provide a preparation method and application of Mongolian medicine tribulus terrestris ethanol extract, which aims to solve the problems in the prior art and provide a novel substance for preparing hypoglycemic drugs.
In order to achieve the purpose, the invention provides the following scheme:
one of the technical schemes provides a preparation method of a caltrop ethanol extract, which comprises the following steps: soaking fructus Tribuli powder for 2h, adding 10 times of 95% ethanol, boiling with strong fire, decocting with slow fire for 1h, and collecting supernatant; adding 95% ethanol 8 times the amount of the residue, boiling with strong fire, decocting with slow fire for 1 hr, and collecting supernatant; adding 95% ethanol 6 times the amount of the residue, boiling with strong fire, decocting with slow fire for 1 hr, and collecting supernatant; mixing the supernatants for 3 times, and concentrating under reflux.
The second technical proposal provides a caltrop ethanol extract prepared by the preparation method.
The third technical proposal provides the application of the tribulus terrestris ethanol extract in preparing hypoglycemic drugs.
The fourth technical proposal provides the application of the caltrop ethanol extract in preparing antioxidant drugs.
The fifth technical scheme provides the application of the tribulus terrestris ethanol extract in preparing the medicine for resisting the hepatocyte damage.
The sixth technical scheme provides an application of a group of serum biomarkers as a detected object in screening hypoglycemic active ingredients of a caltrop ethanol extract, wherein the biomarkers comprise the following components: cytosine, betaine, nicotinamide, piperic acid, piperidine-1-carboxylic acid, L- (+) -lysine, pyridoxamine, cysteine, indoleacetic acid, trimethyllysine, S n-glycerol-3-phosphocholine, pyrrole-2-carboxylic acid, taurine, b-guanidinopropionic acid ester.
The invention discloses the following technical effects:
the invention provides effective data and experimental results of the hypoglycemic effect of the tribulus terrestris ethanol extract. According to the invention, the effect and possible molecular mechanism of the tribulus terrestris ethanol extract for reducing hyperglycemia induced by high-fat feed by regulating lipid metabolism are observed through a rat hyperglycemia model induced by the high-fat feed. The results show that, for hyperglycemia induced by high-fat feed, the ethanol extract of the tribulus terrestris can improve the indexes of metabolic components to be close to normal values, improve the indexes of antioxidase SOD, CAT and GSH-Px, reduce the index of serum MDA and reduce the apoptosis rate of liver cells. The hypoglycemic effect is expressed by the effects of resisting oxidation, reducing the activation of the apoptosis rate channel of the liver cells and the like.
The 18 biomarkers related to the caltrop ethanol extract provided by the invention can play a role in the subsequent separation, research and identification of effective components in the caltrop ethanol extract.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings needed in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings without creative efforts.
FIG. 1 is a workflow diagram of metabolic profiling;
FIG. 2 is a QC positive ion chromatogram;
FIG. 3 is a QC anion chromatogram;
FIG. 4 is a PCA score plot for quality control sample evaluation;
FIG. 5 is a graph of a PCA analysis of a high dose group (HD) versus a model group (model); HD on the left and model on the right in the figure; in four points at the junction, the model group is positioned at the lower right part, and the rest are HD groups;
FIG. 6 is a plot of the PLS-DA analysis for the high dose group (HD) versus the model group (model); HD on the left and model on the right in the figure;
FIG. 7 is a graph of an analysis of OPLS-DA in the high dose group (HD) versus the model group (model); HD on the left and model on the right in the figure;
FIG. 8 is a thermographic analysis of the top 25T-test ranking of the high dose group (HD) compared to the model group (model);
FIG. 9 is a volcanic image; the upper left and upper right regions of the plot are spots of greater than 1.5 in the fold test and less than 0.05 in the T test;
FIG. 10 is a diagram of metabolic pathway analysis; each point in the graph represents a path;
FIG. 11 is a graph of differential metabolite metabolism pathway impact factor analysis between the high dose group (HD) and the model group (model);
FIG. 12 is an index map of antioxidant activity of ethanol extract of Tribulus terrestris;
FIG. 13 is a graph showing the inhibition of hepatocyte apoptosis rate by ethanol extract of Tribulus terrestris; in the figure, (A) is Normal group; (B) the cells are in MODEL group; (C) is in HD group; (D) is MD group; (E) is set as LD.
Detailed Description
Reference will now be made in detail to various exemplary embodiments of the invention, the detailed description should not be construed as limiting the invention but as a more detailed description of certain aspects, features and embodiments of the invention.
It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. The materials, instruments, reagents and experimental methods used in the present invention are all the materials, instruments, reagents and experimental methods which are conventional in the art unless otherwise specified.
It will be apparent to those skilled in the art that various modifications and variations can be made in the specific embodiments of the present disclosure without departing from the scope or spirit of the disclosure. Other embodiments will be apparent to those skilled in the art from consideration of the specification. The specification and examples are exemplary only.
Example 1
Effect of Tribulus terrestris ethanol extract on in vivo metabolism of hyperglycemic rats
1. Materials and methods
1.1 Experimental materials, reagents and instruments
Experimental animals: male Wistar rats of SPF class 100, body weight (200 ± 20) g, purchased from longliving creatures gmbh, No.: SCXK (Liao) 2015-0001. Feeding in water at a free time, and keeping the environment at a temperature of 22 + -1 deg.C and a relative humidity of 45 + -5% for 12 h/d.
Experimental drugs: tribulus terrestris, provided by the formulation room of the hospital affiliated with the university of inner Mongolia, water (Water, Dreches); mass spectrometry acetonitrile (Fisher); formic acid (Sigma-Aldrich). D-galactose, CAT, SOD, GSH-Px, MDA kit (Roche diagnostics GmbH, Germany).
The main experimental apparatus: vortex oscillator (vortex), M16710-33, Thermo Scientific; ultrasonic cleaner (Ultrasonic Washing Device), KQ5200DE, Ultrasonic instruments ltd, kunshan; a bench high speed refrigerated centrifuge (high speed refrigerated centrifuge), hdcroflux 22R, Beckman Coulter; mass spectrometer (MASS), tripleTOF5600+, AB SCIEXTM(ii) a Chromatography (UHPLC), Nexera UHPLC LC-30A, Shimadzu; RE52CS rotary evaporator (Shanghai Yangrong Biochemical instruments factory), DZTW type temperature-adjusting electric heating jacket, cobas c 311 full-automatic biochemical instrument, and New Brunswick ultra-low temperature refrigerator.
1.2 Experimental methods
1.2.1 preparation of ethanol extract of Tribulus terrestris
2kg of raw caltrops are taken, fried to be light yellow by mild fire, and ground by a grinder after smelling the fragrance to prepare the water extract of the caltrops. The ethanol extraction method comprises the following steps: soaking appropriate amount of fructus Tribuli powder for 2h, placing in an electric heating jacket, adding 10 times of 95% ethanol, boiling with strong fire, decocting with slow fire for 1h, and collecting supernatant; adding 95% ethanol 8 times the amount of the residue, boiling with strong fire, decocting with slow fire for 1 hr, and collecting supernatant; adding 95% ethanol 6 times the amount of the residue, boiling with strong fire, decocting with slow fire for 1 hr, and collecting supernatant; the supernatant was combined 3 times and concentrated to a 1mg/ml solution by rotary evaporator reflux for further use.
1.2.2 treatment of laboratory animals
Rats were randomly divided into 5 groups of 10 rats each, including a normal control group, a MODEL control group (MODEL), and three dose groups of High (HD), Medium (MD) and Low (LD) alcohol extract of Tribulus terrestris. After adaptive feeding for 1 week, each model group rat was fed with high-fat diet 42d, and after the model was successful, the rats were gavaged with the corresponding dose of drug solution (0.0073g/mL, 0.0147g/mL, 0.0293g/mL) 1 time per day for 14 d. After the last administration, the rats in each group were anesthetized, blood was taken from the abdominal aorta, the blood glucose content was measured, the remaining samples were centrifuged at 12000r/min for 10min, and serum was separated and stored at-80 ℃ for further use.
Thawing the sample at 4 deg.C, adding 200 μ L serum into 600 μ L cold acetonitrile, and performing ultrasonic treatment in ice bath for 30min 1; centrifuging at 2000rpm and 4 deg.C for 10min, and vacuum centrifuging the supernatant at 37 deg.C to dry. Dissolving the residue with 100 μ L acetonitrile, centrifuging at 12000rpm and 4 deg.C for 10min, sampling supernatant 10 μ L, and UHPLC-MS detecting.
1.2.3 data analysis and processing
Metabolic profile analysis based on mass spectrometry was performed to look for differential biomarkers in different groups and KEGG analysis was performed to study their pathways. The metabolic profile analysis flowchart is shown in FIG. 1.
2. Results of the experiment
The effect of the ethanol extract of tribulus terrestris on blood glucose in hyperglycemic rats is shown in table 1. The results show that the ethanol extract of the tribulus terrestris can reduce the blood sugar of the rat with hyperglycemia.
TABLE 1 ethanol extract of Tribulus terrestris on hyperglycemia rat blood glucose
The chromatogram is shown in fig. 2 and 3. Quality Control (QC) is usually required during the analysis. Theoretically, QC samples are the same, but systematic errors exist in the sample extraction and detection analysis processes, so that differences exist among QC samples, and the smaller the difference is, the higher the stability of the method is, the better the data quality is. The PCA score plot is shown in FIG. 4, which shows a dense distribution of QC samples and reliable data.
Metabonomic data analysis (model group compared to group of high dose quinoa ethanol extract): group HD and group MODEL samples were analyzed and PCA analysis by unsupervised technique (FIG. 5), PLS-DA analysis by supervised technique (FIG. 6) and OPLS-DA analysis by supervised technique (FIG. 7), thermographic analysis (FIG. 8), and volcanic chart analysis (FIG. 9), respectively. In the following analysis, the interpretation ratio of the principal component is shown in parentheses (xx.x%), for example, and the interpretation ratio of the model can be obtained by adding the principal components of PC1 and PC 2. The circles represent 95% confidence intervals.
The PCA analysis is shown in FIG. 5. And analyzing the group HD and group MODEL samples, and respectively performing PCA analysis of an unsupervised technology, PLS-DA analysis of a supervised technology and OPLS-DA analysis, heat map analysis and volcanic chart analysis.
PLS-DA analysis is shown in FIG. 6, and from the PLS-DA score plot, it can be seen that HD and MODEL can be completely separated, so that the MODEL can be used as a separation MODEL for separating two samples. The model is subjected to parameter verification. R2X represents the interpretation ratio for X, i.e. for the principal component in metabolic brackets, e.g. (xx.x%), and adding the principal components of PC1 and PC2 gives the model interpretation ratio. The circles represent 95% confidence intervals. The prediction rate of metabolites R2X is negligible in this model. R2Y represents the prediction rate for Y, i.e., the grouping, indicating that the prediction rate of the model is above 70%, and Q2 represents the accuracy of the model prediction, as shown in the following table:
TABLE 2PLS-DA model parameter testing
| Measure | 1comps | 2comps | 3comps | 4comps | 5comps |
| Accuracy | 0.95 | 0.95 | 0.95 | 0.95 | 0.95 |
| R2 | 0.81171 | 0.96074 | 0.98341 | 0.99721 | 0.99937 |
| Q2 | 0.73548 | 0.75492 | 0.79119 | 0.79599 | 0.78426 |
The OPLS-DA analysis diagram is shown in FIG. 7, and it can be seen from the OPLS-DA score diagram that HD and MODEL can be completely separated, so that the MODEL can also be used as a separation MODEL for separating two samples. The model is then subjected to parametric inspection. The prediction rate of the model reaches 84.1%, and Q2 represents the accuracy rate of model prediction, wherein Q2 is 74.7% in the model. To prevent false positives for the model, the model was therefore tested with 100 permutation of response tests, resulting in R2 close to 1 and Q2 of 79.1%.
The heat map analysis is shown in figure 8: thermographic analysis of material before T-test alignment 25.
Volcano plot analysis as shown in fig. 9, using log2 (fold test) as the abscissa and-log 10(T test) as the ordinate, a volcano plot was obtained with red dots of greater than 1.5 in the fold test and less than 0.05 in the T test.
The generation of hyperglycemia is probably related to the oxidative stress in rats and the activation of hepatocyte apoptosis metabolic pathway, and the ethanol extract of the caltrop can intervene the related pathway, thereby achieving the purpose of treating hyperglycemia. The results of the pathway enrichment analysis for all the detected substances screened are shown in FIG. 10. Rats (rat) were selected as background for enrichment analysis based on the ID of all metabolites in the KEGG database. The enriched pathways are plotted using an abscissa approximation (approximation is the path approximated value from path topology analysis), and an ordinate-log (p). The red boxes represent the detected substance, the blue boxes represent the undetected substance in the pathway, and the number in the box is the KEGG number for that substance. It is shown to be involved in sugar metabolism, amino acid metabolism, fat metabolism, tricarboxylic acid cycle and other pathways of the body.
Differential metabolite metabolic pathways were analyzed by the MetPA database, which is part of the metaboanalyst (www.metaboanalyst.ca), based mainly on KEGG metabolic pathway analysis. The MetPA database identifies possible metabolic pathways disturbed by organisms through metabolic pathway concentration and topological analysis, and then analyzes the metabolic pathways of the metabolites, and an HD vs MODEL-differential metabolite metabolic pathway influence factor analysis chart is shown in FIG. 11.
And (4) combining mass spectrum information to perform systematic identification and identification on the differential metabolites. 18 differential biomarkers were identified, with specific substances as shown in table 3:
TABLE 318 differential biomarkers
The small molecular difference taurine, lysine and the like are all related to oxidative stress metabolic pathways, and the suggestion is that diabetes is possibly caused by metabolic disorder of the oxidative stress metabolic pathways. The small molecule differential cysteine is associated with the apoptotic pathway of hepatocytes, homocysteine promotes hepatocyte apoptosis, and MST1(mammalian mutant 20-like kinase1) may play a role in this process. Suggesting that diabetes may be caused by metabolic dysregulation of the apoptotic stress pathway in hepatocytes.
Choline compounds are the main components of cell membranes, and the decomposition of fat by choline compounds produces a large amount of free fatty acids, which may affect insulin secretion, and at the same time, excess choline generates acetylcholine by combining with acetyl-coa, which inhibits the tricarboxylic acid cycle. Compared with the normal group, the Sn-glycerol-3-phosphorylcholine level of the model group is obviously increased, the fat in the body of the model rat is increased, more choline metabolic cholesterol is generated, the free fatty acid level is increased, and the insulin secretion is possibly inhibited.
Lysine is one of the structural components of the molecule of glycosylation endproducts, which is formed from sugar oxidation and lipid peroxidation. The level of L- (+) -lysine in the experimental model group is obviously increased, and the level is obviously reduced after the ethanol extract of the caltrops is dried. Therefore, the ethanol extract of the tribulus terrestris participates in the processes of sugar oxidation and lipid peroxidation in the rat body of the hyperglycemia model.
Plasma betaines are associated with human insulin sensitivity. Betaine supplementation in high fat fed mice improved glucose homeostasis, liver fat, insulin sensitivity, and increased energy expenditure and oxidative capacity in the white adipose tissue of the groin. The betaine level in the model group is obviously increased compared with that in the normal group, and the level is reduced after the ethanol extract of the caltrops is dried. Therefore, the ethanol extract of the tribulus terrestris participates in the processes of sugar oxidation and lipid peroxidation in the rat body of the hyperglycemia model.
Nicotinamide is related to promotion of fat cell energy metabolism, and various fat energy metabolism related diseases such as obesity and fatty liver are related to indexes of nicotinamide. Compared with the normal group, the nicotinamide level of the model group is obviously increased, and the level is reduced after the ethanol extract of the caltrops is dried. Therefore, the ethanol extract of the tribulus terrestris participates in the processes of sugar oxidation and lipid peroxidation in the rat body by intervening in the fat metabolism of the rat in a hyperglycemia model.
Piperic acid is also involved in fat metabolism. Compared with the normal group, the model group has significantly increased piperic acid level, and the level is reduced after the ethanol extract of the caltrops is dried. Therefore, the ethanol extract of the tribulus terrestris participates in the processes of sugar oxidation and lipid peroxidation in the rat body by intervening in the fat metabolism of the rat in a hyperglycemia model.
The piperidine-1-formic acid is one of in vivo protein metabolites, and compared with a normal group, the piperidine-1-formic acid level in a model group is obviously increased, and the piperidine-1-formic acid level in a dry state after the ethanol extract of the tribulus terrestris is obviously reduced. Therefore, the ethanol extract of the tribulus terrestris interferes with the index of the poor metabolism foreign body piperidine-1-formic acid.
The b-guanidinopropionic acid lipid index is related to the improvement of insulin sensitivity and the selective reduction of adipose tissue weight, and the b-guanidinopropionic acid can treat and prevent metabolic disorder, hyperglycemia, hyperinsulinemia, insensitivity of blood insulin, hyperlipidemia, blood amylase excess, obesity and other symptoms. Compared with the normal group, the level of b-guanidinopropionate in the model group is obviously reduced, and the level is obviously increased after the ethanol extract of the tribulus terrestris is dried.
Lysine is one of the structural components of the molecule of glycosylation endproducts, which is formed from sugar oxidation and lipid peroxidation. The level of L- (+) -lysine in the model group is obviously increased, and the level is obviously reduced after the ethanol extract of the caltrops is dried. Therefore, the ethanol extract of the tribulus terrestris participates in the processes of sugar oxidation and lipid peroxidation in the rat body of the hyperglycemia model.
Cysteine is a metabolite generated by conversion of methionine in proteins, is an intermediate product of methionine metabolized into glutathione (the most important antioxidant enzyme in vivo) and S-adenosylmethionine (SAMe), and can promote hepatocyte apoptosis. Elevated plasma cysteine levels are an independent risk factor for cardiovascular and cerebrovascular diseases. Compared with the normal group, the cysteine level of the model group is obviously increased, and the level is obviously reduced after the ethanol extract of the caltrops is dried. Therefore, the ethanol extract of the tribulus terrestris has the function of inhibiting the cysteine level.
Example 2
Antioxidation identification of caltrop ethanol extract
SOD, MDA, CAT, GSH-Px of mice of different groups were detected, and the results are shown in FIG. 12. SOD, MDA, CAT, GSH-Px on oxidative stress pathway are changed, and the drug has the function of adjusting the change. The experimental results are as follows: the indexes of CAT, serum SOD and GSH-Px in blood of a model group model rat are all obviously reduced, and the index of MDA in serum is obviously increased. However, the index of the rat gavaged with the Tribulus terrestris extract tends to return to normal value. Has statistical significance (P is less than 0.01).
Example 3
Fructus Tribuli ethanol extract for inhibiting hepatocyte apoptosis
The effect of aqueous extracts of tribulus terrestris on apoptosis in model rat hepatocytes is shown in figure 13. Compared with the normal control group, the number of the hepatocyte apoptosis of the model group is obviously increased, and the statistical significance is achieved (P is less than 0.01). Compared with the model group, the hepatic cell apoptosis number of the group with three doses of large, medium and small caltrops is obviously reduced, and the statistical significance is achieved (P is less than 0.01). The tribulus terrestris aqueous extract has the function of inhibiting the apoptosis of model liver cells.
The above-described embodiments are merely illustrative of the preferred embodiments of the present invention, and do not limit the scope of the present invention, and various modifications and improvements of the technical solutions of the present invention can be made by those skilled in the art without departing from the spirit of the present invention, and the technical solutions of the present invention are within the scope of the present invention defined by the claims.
Claims (6)
1. A preparation method of a caltrop ethanol extract is characterized by comprising the following steps: soaking fructus Tribuli powder for 2h, adding 10 times of 95% ethanol, boiling with strong fire, decocting with slow fire for 1h, and collecting supernatant; adding 95% ethanol 8 times the amount of the residue, boiling with strong fire, decocting with slow fire for 1 hr, and collecting supernatant; adding 95% ethanol 6 times the amount of the residue, boiling with strong fire, decocting with slow fire for 1 hr, and collecting supernatant; mixing the supernatants for 3 times, and concentrating under reflux.
2. An ethanol extract of tribulus terrestris prepared according to the preparation method of claim 1.
3. Use of the ethanol extract of tribulus terrestris as claimed in claim 2 in the preparation of hypoglycemic drugs.
4. An application of the ethanol extract of tribulus terrestris as claimed in claim 2 in preparing antioxidant drugs.
5. Use of the ethanol extract of tribulus terrestris of claim 2 in the preparation of a medicament for treating hepatocyte injury.
6. The application of a group of serum biomarkers as a detected object in screening hypoglycemic active ingredients of ethanol extract of tribulus terrestris is characterized in that the biomarkers comprise the following components: cytosine, betaine, nicotinamide, piperic acid, piperidine-1-carboxylic acid, L- (+) -lysine, pyridoxamine, cysteine, indoleacetic acid, trimethyllysine, Sn-glycerol-3-phosphorylcholine, pyrrole-2-carboxylic acid, taurine, b-guanidinopropionic acid ester.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110529731.5A CN113134020A (en) | 2021-05-14 | 2021-05-14 | Preparation method and application of Mongolian medicine tribulus terrestris ethanol extract |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110529731.5A CN113134020A (en) | 2021-05-14 | 2021-05-14 | Preparation method and application of Mongolian medicine tribulus terrestris ethanol extract |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN113134020A true CN113134020A (en) | 2021-07-20 |
Family
ID=76817391
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110529731.5A Pending CN113134020A (en) | 2021-05-14 | 2021-05-14 | Preparation method and application of Mongolian medicine tribulus terrestris ethanol extract |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113134020A (en) |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101234133A (en) * | 2007-11-09 | 2008-08-06 | 西安天健医药科学研究所 | Technique for using tribulus whole grass to extract total saponins of tribulus |
| CN101259153A (en) * | 2008-04-25 | 2008-09-10 | 江西远健药业有限公司 | Tribulus total steroid saponin and preparation thereof |
| CN108938687A (en) * | 2018-09-03 | 2018-12-07 | 辽宁何氏医学院 | Puncture vine prevents and treats the application in retinal damage disease medicament in preparation |
| CN109142419A (en) * | 2018-09-30 | 2019-01-04 | 广西医科大学 | A kind of potential marker metabolic pathway of serum and research method of the moringa seeds anti-diabetic based on metabolism group |
| CN109419826A (en) * | 2017-08-30 | 2019-03-05 | 南京泽朗医药技术有限公司 | A kind of preparation method of Fructus Tribuli crude saponin |
-
2021
- 2021-05-14 CN CN202110529731.5A patent/CN113134020A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101234133A (en) * | 2007-11-09 | 2008-08-06 | 西安天健医药科学研究所 | Technique for using tribulus whole grass to extract total saponins of tribulus |
| CN101259153A (en) * | 2008-04-25 | 2008-09-10 | 江西远健药业有限公司 | Tribulus total steroid saponin and preparation thereof |
| CN109419826A (en) * | 2017-08-30 | 2019-03-05 | 南京泽朗医药技术有限公司 | A kind of preparation method of Fructus Tribuli crude saponin |
| CN108938687A (en) * | 2018-09-03 | 2018-12-07 | 辽宁何氏医学院 | Puncture vine prevents and treats the application in retinal damage disease medicament in preparation |
| CN109142419A (en) * | 2018-09-30 | 2019-01-04 | 广西医科大学 | A kind of potential marker metabolic pathway of serum and research method of the moringa seeds anti-diabetic based on metabolism group |
Non-Patent Citations (2)
| Title |
|---|
| 崔佳乐等: "蒺藜的药效学探讨", 《齐齐哈尔医学院学报》 * |
| 王胜勇等: "《中药高清原大图谱》", 31 January 2020, 福建科学技术出版社 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Jacob et al. | A targeted metabolomics approach for clinical diagnosis of inborn errors of metabolism | |
| Ibáñez et al. | Metabolomics, peptidomics and proteomics applications of capillary electrophoresis-mass spectrometry in Foodomics: A review | |
| Wang et al. | Thyroxine and reserpine-induced changes in metabolic profiles of rat urine and the therapeutic effect of Liu Wei Di Huang Wan detected by UPLC-HDMS | |
| Li et al. | Plasma metabolic profiling of dairy cows affected with clinical ketosis using LC/MS technology | |
| Lustgarten et al. | Serum predictors of percent lean mass in young adults | |
| Sun et al. | Exploring potential biomarkers of coronary heart disease treated by Jing Zhi Guan Xin Pian using high-throughput metabolomics | |
| Shan et al. | Untargeted metabolomics reveal the therapeutic effects of Ermiao wan categorized formulas on rats with hyperuricemia | |
| Zheng et al. | Protective effect of cultured bear bile powder against dimethylnitrosamine-induced hepatic fibrosis in rats | |
| Fu et al. | An integrated study on the comprehensive mechanism of Schisandra chinensis polysaccharides mitigating Alzheimer's disease in rats using a UPLC-Q-TOF-MS based serum and urine metabolomics strategy | |
| Li et al. | Correlation between gut bacteria Phascolarctobacterium and exogenous metabolite α-linolenic acid in T2DM: a case-control study | |
| Zhang et al. | Shenqi compound ameliorates type-2 diabetes mellitus by modulating the gut microbiota and metabolites | |
| Yan et al. | Label-free mass spectrometry-based proteomics for investigating the therapeutic mechanism of Sijunzi decoction on spleen deficiency syndrome | |
| Ren et al. | Identification of the perturbed metabolic pathways associating with renal fibrosis and evaluating metabolome changes of pretreatment with Astragalus polysaccharide through liquid chromatography quadrupole time-of-flight mass spectrometry | |
| Nie et al. | Comparison of two types of vinegar with different aging times by NMR‐based metabolomic approach | |
| Jiang et al. | Revealing the hypoglycemic effects and mechanism of GABA-rich germinated Adzuki beans on T2DM mice by untargeted serum metabolomics | |
| Ma et al. | Therapeutic mechanisms of mulberry leaves in type 2 diabetes based on metabolomics | |
| Zhang et al. | Emerging applications of metabolomics to assess the efficacy of traditional Chinese medicines for treating type 2 diabetes mellitus | |
| Pan et al. | Urinary metabolomics study of the intervention effect of hypoglycemic decoction on type 2 diabetes mellitus rats model | |
| An et al. | Metabolomics approach to identify therapeutically potential biomarkers of the Zhi-Zi-Da-Huang decoction effect on the hepatoprotective mechanism | |
| Shi et al. | Comparative pharmacokinetic study of the components of Jia‐Wei‐Kai‐Xin‐San in normal and vascular dementia rats by ultra‐fast liquid chromatography coupled with tandem mass spectrometry | |
| Li et al. | Tian-huang formula, a traditional chinese medicinal prescription, improves hepatosteatosis and glucose intolerance targeting akt-srebp nexus in diet-induced obese rats | |
| Chen et al. | Metabolomics study of Angelica sinensis (Oliv.) Diels on the abnormal uterine bleeding rats by ultra‐performance liquid chromatography–quadrupole–time‐of‐flight mass spectrometry analysis | |
| He et al. | Astragali Radix–Codonopsis Radix–Jujubae Fructus water extracts ameliorate exercise‐induced fatigue in mice via modulating gut microbiota and its metabolites | |
| Dong et al. | In vivo anti-hyperuricemia and anti-gouty arthritis effects of the ethanol extract from Amomumvillosum Lour. | |
| CN113134020A (en) | Preparation method and application of Mongolian medicine tribulus terrestris ethanol extract |
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 | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20210720 |
|
| RJ01 | Rejection of invention patent application after publication |