| Heart failure is the terminal stage of a variety of cardiovascular diseases.Myocardial infarction is one of the common causes of heart failure.Although the treatment of myocardial infarction has made advanced breakthrough,patients with myocardial infarction are still at risk of heart failure.The evolution from myocardial infarction to heart failure is a dynamic,complex and chain reaction,including the loss of myocardial cells,neurohumoral system activation,heart remodeling,etc.Static observation of myocardial infarction and heart failure restricts the analysis of the evolution of these disease.This study focuses on the dynamic changes of the evolution process of myocardial infarction induced heart failure.By analyzing the phenotype and molecular changes of myocardial infarction induced heart failure,it is helpful to analyze the evolution process from myocardial infarction to heart failure,reveal the interaction rules between different time and different links,and lay the foundation for elucidating the molecular mechanism of myocardial infarction induced heart failure.It is difficult to recover once heart failure occurs,"early detection and early treatment" are essential in the treatment of heart failure.Biomarkers are helpful for clinical diagnosis and prognosis of patients with heart failure.Although the research on biomarkers of heart failure has made some progress,the clinical application of new biomarkers is limited due to limited detection,unknown reaction to drugs and easily affected by non cardiogenic factors;In addition,the current biomarkers mainly focus on the diagnosis and prognosis of heart failure,lack of markers to provide early warning information,so it is of great significance to find biomarkers that can play the role of early warning.Traditional Chinese medicine is widely used in clinical treatment because of its good curative effect and small toxic and side effects.However,its complex components and effects on complex organisms make it more difficult to analyze the mechanism of action and main intervention links of traditional Chinese medicine.Shengmai Injection(SMI)is derived from Shengmai Powder.It is recommended for acute,chronic and refractory heart failure because of its simple composition and good curative effect.However,the present research of Shengmai injection is mostly focused on clinical observation,and there are few studies on the main action links of heart failure,especially myocardial infarction.Therefore,it is very important to carry out the research on the mechanism of action and the main action links of Shengmai injection..To sum up,in view of the problems of unknown pathogenesis,lack of early warning markers,and unclear intervention links of Shengmai Injection in the evolution process of myocardial infarction to heart failure,this project first studied the evolution process of myocardial infarction induced heart failure,and then screened and verified the potential early warning markers of myocardial infarction induced heart failure,Finally,according to the dynamic differential proteins of myocardial infarction induced heart failure,the main intervention links of Shengmai injection were studied and verified.This study aims to serve the clinical,find potential early warning markers,clarify the main intervention links of Shengmai injection,and provide research strategies for the clinical precise application of Shengmai injection.Content1.The section one focused on the evolution of heart failure caused by myocardial infarction.The blood biochemical indexes,cardiac function and pathological changes from 1 week to 4 weeks after myocardial infarction were analyzed by ligation model of left coronary artery anterior descending branch in rats;Then,the differently expressed proteins of left ventricular at different time point from myocardial infarction to heart failure was analyzed by proteomics,and enrichment annotion was performed using bioinformaticsinformation biology.The evolution of heart failure caused by myocardial infarction was revealed by combining the disease phenotype and molecular characteristics.2.The section two focused on the analysis and verification of the potential early warning markers of heart failure caused by myocardial infarction.Firstly,the differently expressed proteins were analyzed,and the candidate markers with early warning function were screened;Secondly,the level of candidate markers in serum of rats with heart failure caused by myocardial infarction was detected by ELISA;Finally,the selected markers were verified at the cell level.After treatment with AngⅡ,the level of candidate markers was analyzed by immunofluorescence method,and the changes of potential markers after the Shengmai injection intervention were analyzed by immunofluorescence method.3.The section three focused on the main intervention links of Shengmai injection analysis.The main intervention of Shengmai injection were predicted and analyzed by network pharmacology,and the main intervention effect of Shengmai injection was confirmed from animal level and cell level.The animal level was performed with the ligation model of left coronary artery anterior descending branch in rats,and the pharmacodynamic evaluation of Shengmai injection was conducted from the heart function and pathological results;The expression levels of VCAN,COL1A1 and P ⅠCP were detected by ELISA after the intervention of Shengmai injection.After Shengmai Injection treatment,the LDH,ROS and the level of Ca2+were detected and the level of ANP and CTGF were analyzed by immunofluorescence.ResultsSection one:the study of the evolution from myocardial infarction to heart failure1.Changes in disease phenotypeIn the dynamic evolution process from myocardial infarction to heart failure,abnormalities of cardiac function occurred firstly.The ejection fraction and short axis shortening rate of the model group decreased significantly in the first week after ligation,and in the second week,the 3rd and 4th week continued to decrease,indicating that the myocardial contractility was impaired;the left heart weight ratio to the heart weight ratio increased significantly at second week,and continue increase at the 3rd and 4th weeks,indicating that myocardial hypertrophy occurred at second week;compared with sham group,rats occured myocardial tissue structure abnormality and myocardial fibrosis injury at 3rd and 4th week in model group;the level of BNP and NT-proBNP significantly increased at the 4th week in model group,indicating that the ventricular load was overloaded.2.Proteomics results(1)A total of 6127 proteins were identified through proteomics analysis.Compared with sham group,there were 838 differentially expressed proteins at first week in model group,716 differentially expressed proteins at 2nd week in model group,416 differentially expressed proteins at 3rd week in model group,and 781 differentially expressed proteins at 4th week in model group.There are a large number of proteins up-regulated at the later stages of the evolution from myocardial infarction to heart failure.(2)From week 1 to week 4,differentially expressed proteins are involved in calcium ion transport-related biological functions,oxidative stress,cardiac contraction,heart rate regulation and other functions;proteins differentially expressed at week 2 are also involved in inflammation,apoptosis,etc.Differentially expressed proteins related to myofibril production appeared at the 3rd week,and the differentially expressed proteins also participated in the production of extracellular matrix at the fourth week.Section two:Discovery and verification of early warning biomarkers for m yocardial infarction induced heart failure caused1.Screening of potential early warning biomarkersBased on the unsupervised model,a total of 33 proteins that was related to disease typing have early warning significance during the evolution from myocardial infarction to heart failure were screened out;COX single factor and multivariate regression analysis were used to screen out candidate markers.2.Animal level verification resultsThe ELISA was used to detect the expression levels of VCAN and COL1A1 in rat serum,and it was found that the expression levels of VCAN and COL1A1 increased significantly at 3rd and 2nd weeks respectively(p<0.05),and still significantly increased at 4th week(p<0.05).3.Cell level verification results(1)Using immunofluorescence to detect the expression level of VCAN and COL1A1 after 10-4 μM to 10-6 μM Ang Ⅱ treated H9c2 cells for 24 h.The expression level of VCAN was found to be significant increased in the 10-3 μM and 10-4 μM in Ang Ⅱ treatment groups(p<0.05),COL1A1 was significantly increased after 10-4μM to 10-5 μM Ang Ⅱ treatment(p<0.05).(2)After intervention with Shengmai injection(1 μL/4 mL to 1 μL/2 mL),the expression level of COL1A1 decreased significantly(p<0.05).Section three:Investigation of main intervention of Shengmai injection1.The results of prediction analysis of the precise intervention of Shengmai injectionA total of 190 chemical components and 1,612 targets of SMI were collected to construct a "component-target" network;Shengmai injection is mainly involved in calcium channel regulation,inflammation,myocardial contraction,oxidative stress,cardiac Biological functions such as muscle fibrosis.Through the protein-protein analysis,it is found that the target spectrum of Shengmai injection is significantly related to the first week,second week,and third week after myocardial infarction.At 2nd week,the number of targets established correlation was the highest.After enrichment annotion,alleviation myocardial fibrosis may play an important role of SMI during the period from myocardial infarction to heart failure.2.Validation results of the precise intervention of Shengmai injectionAt the animal level,Shengmai injection can improve heart function,reduce myocardial fibrosis injury,and the expression level of VCAN,COL1A1 and PICP;at the cellular level,Shengmai injection can slow down the degree of cell damage and reduce the excessive accumulation of ROS and level of Ca2+,and the expression of level of CTGF and ANP.Conclusion1.The results showed that the cardiac function gradually decreased,myocardial hypertrophy,myocardial fibrosis and ventricular mechanical stress gradually increased in the process of myocardial infarction to heart failure;Proteomic analysis showed that myocardial infarction induced heart failure was mainly related to extracellular matrix formation,inflammation and other biological processes.2.It is confirmed that the changes of Vcan and COL1A1 are related to myocardial infarction induced heart failure,which may provide reference information for the development and prognosis of heart failure.3.Network pharmacology technology was used to analyze the target of Shengmai injection and the dynamic molecular characteristic spectrum in the process of myocardial infarction induced heart failure.The results showed that the main intervention link of Shengmai Injection in the process of myocardial infarction induced heart failure may be related to reducing myocardial fibrosis damage;Animal experiment confirmed that Shengmai injection can slow down myocardial fibrosis injury caused by myocardial infarction,and cell experiment confirmed that Shengmai injection can reduce myocardial cell injury caused by Ang Ⅱ.This study shows that it is feasible to analyze the main intervention links of listed Chinese patent medicines in the evolution process from myocardial infarction to heart failure based on the dynamic molecular characteristic spectrum of myocardial infarction induced heart failure.The innovation point of the study1.The evolution from myocardial infarction to heart failure includes many biological processes,such as the loss of myocardial cells,the activation of neurohumoral system and so on.This study focuses on the dynamic changes of the evolution process of myocardial infarction induced heart failure.By analyzing the phenotype and molecular changes from myocardial infarction to heart failure,we can reveal the dynamic evolution process of myocardial infarction induced heart failure,and on this basis,we can more accurately screen the early warning markers of myocardial infarction induced heart failure;2.It is suggested that COL1A1 and Vcan may be potential early warning markers of myocardial infarction induced heart failure;3.Taking Shengmai injection as an example,this study adopts the analysis strategy of "dynamic molecular characteristic spectrum-main intervention links-experimental verification" to predict and verify the main intervention links of Shengmai injection.This study will provide research strategies for the precise application of other listed Chinese patent medicines in the process of myocardial infarction to heart failure. |
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