CN-121971578-A - Discovery of anti-heart failure traditional Chinese medicine monomer composition based on Zhenwu decoction, and preparation method and application thereof

Abstract

The invention discloses a traditional Chinese medicine monomer composition for resisting heart failure based on Zhenwu decoction, which comprises the steps of preparing Zhenwu decoction, collecting and analyzing serum samples, determining endogenous metabolites for resisting heart failure and measuring peak intensity, determining absorbed blood components and measuring peak intensity, and analyzing correlation of peak intensity. The invention simultaneously obtains the endogenous metabolite of heart failure resistance and the blood component absorption information of the Zhenwu decoction through one-time liquid chromatography-mass spectrometry analysis, further obtains the active component of heart failure resistance Wu Shangkang, saves time and improves the accuracy of results, and the traditional Chinese medicine monomer composition has stable and controllable quality, has effective heart failure resistance effect and is suitable for preparing heart failure resistance medicines.

Inventors

• LIANG JUNLI
• ZHOU QIAN
• TAN GUANGGUO

Assignees

• 中国人民解放军空军军医大学

Dates

Publication Date

20260505

Application Date

20260212

Claims (5)

1. 1. The discovery of the anti-heart failure traditional Chinese medicine monomer composition based on Zhenwu decoction is characterized in that the discovery process comprises the following steps: Preparation of Zhenwu decoction Mixing radix Aconiti lateralis Preparata, atractylodis rhizoma, poria, radix Paeoniae alba and rhizoma Zingiberis recens at a mass ratio of 3:3:3:2:3, extracting with water, filtering, mixing the extractive solutions, concentrating to obtain ZHENWU decoction, precipitating with ethanol, filtering, concentrating the filtrate under reduced pressure to obtain ZHENWU decoction concentrate; Step two, collecting and analyzing serum samples Step 201, collecting serum sample Dividing male SD rats into three groups, namely a sham operation group, a heart failure model group and a model administration group, wherein the rats of the sham operation group and the heart failure model group are subjected to normal saline lavage every day, the rats of the model administration group are subjected to real wu decoction concentrated solution intragastric administration in the first step, the continuous administration is carried out for 28 days, and after the real wu decoction concentrated solution intragastric administration is carried out for 30 minutes on the 7 th day, the 14 th day, the 21 th day and the 28 th day, blood of each group of rats is taken, and supernatant of each group is obtained after centrifugation and is used as a serum sample to be stored at-80 ℃; step 202, analysis of serum samples Thawing the serum sample in the step 201, respectively adding precooled methanol containing diclofenac and 2-chloro-L-phenylalanine internal standard, swirling, centrifuging, absorbing supernatant, evaporating to dryness, adding methanol water into residues for redissolution and swirling, and then carrying out liquid chromatography-mass spectrometry analysis to obtain data of each group of serum samples; step three, determination of endogenous metabolite of true Wu Shangkang heart failure and determination of peak intensity Step 301, determination of endogenous metabolites of true Wu Shangkang heart failure Processing the data of each group of serum samples on the 28 th day in the data of each group of serum samples in the step 202 after converting the format to obtain a three-dimensional data matrix containing retention time, mass-to-charge ratio and peak intensity, identifying endogenous metabolites through data inquiry, normalizing total peak area and internal standard of peak intensity information to correct mass spectrum response, and screening out 36 abnormal endogenous metabolites of the model group by comparing the changes of the endogenous metabolites of the sham operation group and the heart failure model group with the change multiple of the peak intensity which is more than 1.5 or less than 0.67 and the p value which is less than 0.05; the method comprises the steps of taking the abnormal endogenous metabolites of 36 model groups as potential drug effect indexes, comparing the abnormal endogenous metabolites of 36 heart failure model groups with the abnormal endogenous metabolites of model administration groups, analyzing substances which are obviously reversed in the abnormal endogenous metabolites after the intervention of the Zhenwu decoction by adopting a variance analysis method, and determining 18 kinds of the natural wu Shang Kang heart failure reversed endogenous metabolites; Step 302, determination of peak intensity of endogenous metabolite of true Wu Shangkang heart failure According to the method in step 301, a three-dimensional data matrix containing retention time, mass-to-charge ratio and peak intensity of the model administration group serum samples on the 7 th day, the 14 th day, the 21 st day and the 28 th day in the data of each group serum sample in step 202 is obtained, the endogenous metabolite information of the 18 kinds of Zhenwu Shang Kang heart failure reversals determined in step 301 is extracted from the three-dimensional data matrix, the dynamic change profile of the 18 kinds of Zhenwu Shang Kang heart failure reversals endogenous metabolites from the 7 th day to the 28 th day is obtained through total peak area normalization and internal standard normalization, and the endogenous metabolite peak intensity information of the 18 kinds of Zhenwu Shang Kang heart failure reversals is obtained; Step four, determination of blood absorption component of Zhenwu decoction and determination of peak intensity Step 401, determining blood component absorbed by Zhenwu decoction Diluting the Zhenwu decoction concentrated solution in the first step by using chromatographic methanol, then carrying out vortex mixing, centrifuging, taking supernatant to carry out liquid chromatography-mass spectrometry analysis, constructing a chemical database containing the ion characteristics of Zhenwu decoction components based on literature retrieval, carrying out in-vitro component identification on the Zhenwu decoction by depending on the database, then comparing the peak retention time of each component with the fragment ions, extracting serum migration components of Zhenwu decoction in the rats of the model administration group from the data of each group of serum samples in the step 202, and determining the absorbed blood components of 22 Zhenwu decoction in the rats of the model administration group; Step 402, measuring peak intensity of blood component absorbed by Zhenwu decoction Searching information of absorbed blood components of 22 kinds of Zhenwu decoction in a model administration group rat body in step 401 through retention time and mass-to-charge ratio from a three-dimensional data matrix of serum samples of the model administration group on the 7 th day, the 14 th day, the 21 st day and the 28 th day in step 302, normalizing the total peak area of each serum sample, and then carrying out internal standard normalization to correct mass spectrum response, so as to obtain information of the absorbed blood component peak intensities of the 22 kinds of Zhenwu decoction; Fifthly, analyzing the correlation between the peak intensity of endogenous metabolites of the true Wu Shangkang heart failure and the peak intensity of blood component absorbed by the true Wu decoction And (3) carrying out Pearson correlation analysis on the peak intensity information of the endogenous metabolites of the 18 kinds of Zhenwu Shang Kang heart failure reversals obtained in the step (302) and the peak intensity information of the blood component absorption peaks of the 22 kinds of Zhenwu decoction obtained in the step (402), and using a correlation coefficient r to represent the action degree of the blood component absorption peaks of the Zhenwu decoction on the endogenous metabolites of the Zhen Wu Shangkang heart failure, wherein |r| is more than or equal to 0.6 and p <0.05 represents obvious correlation, thereby screening out the pharmacodynamic substance basis of the Zhen Wu Shangkang heart failure as the component of the traditional Chinese medicine monomer composition.
2. 2. The method according to claim 1, wherein the condition of the liquid chromatography in the step 202 is that a ExionLCTM AD system liquid chromatography system is adopted, a chromatographic column is a Acquity UPLC BEH C column with the specification of 2.1mm×100mm,1.7 μm, a phase A in a mobile phase is formic acid aqueous solution with the formic acid mass content of 0.1%, a phase B is acetonitrile, and gradient elution is adopted, wherein the gradient elution is 0-12 min, and the concentration of B is 5-12%; 12-25 min, 12-20% B, 25-45 min, 20-40% B, 45-55 min, 40-80% B, 55-55.01 min, 80-95% B, 55.01-57 min,95% B, 57-57.01 min, 95-5% B, 57.01-60min,5% B, 0.4mL/min flow rate, 2 mu L sample injection volume, 4 ℃ auto injector temperature, AB SCIEX TripleTOFTM 6600+ mass spectrometer, electrospray ion source (ESI), IDA mode, 5500V and 10V precursor ion scanning range, 40 ℃ column temperature setting, 55psi, 60psi and 35psi ion source gas 1,2 ion source gas and 35psi curtain gas, respectively.
3. 3. The method according to claim 1, wherein the effective substance of Zhen Wu Shangkang heart failure is selected from the group consisting of Song's Guo ling, bighead Atractylodes rhizome lactone II, pachymic acid, paeoniflorin, and 6-gingerol.
4. 4. A method for preparing a traditional Chinese medicine monomer composition based on the findings of any one of claims 1-3, which is characterized in that Song's fruit, atractylenolide II, pachymic acid, paeoniflorin and 6-gingerol are dissolved in a 2% sodium carboxymethyl cellulose solution according to a mass ratio of 9:9:9:6:9, so as to obtain the traditional Chinese medicine monomer composition.
5. 5. The use of a found and obtained traditional Chinese medicine monomer composition according to any one of claims 1-3, wherein the traditional Chinese medicine monomer composition plays a role in preparing an anti-heart failure drug by improving heart function, improving ejection fraction and shortening fraction, improving heart failure myocardial pathological changes, and improving serum functional indexes including creatine kinase CK, creatine kinase isoenzyme CK-MB and lactate dehydrogenase LDH.

Description

Discovery of anti-heart failure traditional Chinese medicine monomer composition based on Zhenwu decoction, and preparation method and application thereof Technical Field The invention belongs to the field of medical biology, and particularly relates to discovery of an anti-heart failure traditional Chinese medicine monomer composition based on Zhenwu decoction, and a preparation method and application thereof. Background Heart failure is a clinical syndrome which is caused by various reasons and has the defects of heart blood pumping function decline and incapacitation of meeting the metabolic needs of organisms, and often relates to various chronic cardiovascular diseases such as coronary heart disease, hypertension, cardiomyopathy and the like, and the heart failure has high morbidity and mortality and seriously threatens the health of human bodies. In general, therapeutic strategies against heart failure include drug therapy, surgical therapy, etc., which are not only costly but also have certain side effects. It is therefore desirable to find an effective drug that can effectively treat heart failure. The natural medicine usually contains various bioactive components, can play a role on a plurality of targets, thereby generating comprehensive and multi-effect treatment effects on diseases, and is more natural due to the fact that the natural medicine is usually extracted from roots, stems, leaves, flowers and other parts of plants, and adverse effects on human bodies are reduced. In recent years, the excavation of effective medicinal ingredients from natural medicines has been attracting attention. The theory of traditional Chinese medicine holds that although heart failure disease is mainly heart, the heart failure disease is closely related to viscera dysfunction such as kidney, small intestine and the like, the pathogenesis of deficiency and excess is basically agreed, and heart qi deficiency and heart yang failure are the principal, phlegm and blood stasis are mutually blocked, and internal retention of water and fluid retention are the principal. The Zhenwu decoction has the efficacy of warming yang and promoting diuresis, consists of aconite, poria cocos, bighead atractylodes rhizome, white paeony root and ginger, and modern researches show that the Zhenwu decoction has obvious heart failure resistance. Although the adverse side effect of the actual Wu Shangkang heart failure is small, the dosage is large, the quality of the decoction can not be ensured to be stable and controllable, and the effect is not obvious like chemical medicines. Therefore, it is particularly important to select an anti-heart failure composition that is found to be more effective from the Zhenwu decoction. Disclosure of Invention The invention aims to solve the technical problem of providing a discovery of a traditional Chinese medicine monomer composition for resisting heart failure aiming at the defects of the prior art. The discovery obtains the heart failure resistant endogenous metabolite of the Zhenwu decoction and the blood component information of the Zhenwu decoction through one-time liquid chromatography-mass spectrometry analysis, combines XCMS package information extraction and analysis of variance to obtain peak intensity information of the two and carries out correlation analysis to obtain the active ingredient of the Zhen Wu Shangkang heart failure, greatly saves the discovery and screening time, improves the accuracy of results, and solves the problem that the prior art lacks to discover an effective heart failure resistant composition from the Zhenwu decoction. In order to solve the technical problems, the invention adopts the technical scheme that the discovery of the traditional Chinese medicine monomer composition for resisting heart failure based on Zhenwu decoction is characterized in that the discovery process comprises the following steps: Preparation of Zhenwu decoction Mixing radix Aconiti lateralis Preparata, atractylodis rhizoma, poria, radix Paeoniae alba and rhizoma Zingiberis recens at a mass ratio of 3:3:3:2:3, extracting with water, filtering, mixing the extractive solutions, concentrating to obtain ZHENWU decoction, precipitating with ethanol, filtering, concentrating the filtrate under reduced pressure to obtain ZHENWU decoction concentrate; Step two, collecting and analyzing serum samples Step 201, collecting serum sample Dividing male SD rats into three groups, namely a sham operation group, a heart failure model group and a model administration group, wherein the rats of the sham operation group and the heart failure model group are subjected to normal saline lavage every day, the rats of the model administration group are subjected to real wu decoction concentrated solution intragastric administration in the first step, the continuous administration is carried out for 28 days, and after the real wu decoction concentrated solution intragastric administration is carried out for 30 minu