The Expression Of NGF And COX-2 In Acute Myocardial Infarction And Its Relationship With TCM Syndromes

According to the latest statistics of the epidemiology,At present,cardiovascular disease is the first death cause in China.Its mortality accounts for more than 40%of the death causes of residents.Especially the incidence of acute myocardial infarction has been increasing rapidly in recent years.It is one of the major diseases that affect people’s health.In the process of occurrence and development of acute myocardial infarction,there is interaction between cardiac remodeling and sympathetic nerve remodeling.It is closely related to the occurrence of malignant arrhythmia and sudden cardiac death after infarction,suggesting a new direction for three level prevention of acute myocardial infarction.Acute myocardial infarction in TCM should belong to the "chest,True heart pain,que heartache" category.There was a corresponding record in the early period of the Huang Di Nei Jing.<Su Wen Q-gas reservoir method on the article>:"heart disease,chest pain,hypochondriac hypochondriac pain over,taking back between two arms between shoulder pain,pain."<LingshuJue>:" really heartache,on foot to the Youth Festival,very heartache,Danfa death,evening once dead." At present,traditional Chinese medicine generally divides the disease into multiple syndromes such as heart blood stasis,phlegm and internal obstruction,but the syndrome type theory does not refer to syndrome Prognosis and risk stratification.It can not provide objective basis for clinical prognosis of all TCM syndromes.This study selected NGF and COX-2 for acute myocardial infarction after sympathetic nerve remodeling markers,through clinical observation and animal experimental cohort as the starting point in order to study the relationship between acute myocardial infarction patients with cardiac sympathetic nerve remodeling and TCM syndrome type.Exploring the law between the acute myocardial infarction patients with different TCM syndromes and the risk level the prognosis and finding TCM regulating cardiac sympathetic nerve remodeling and the theoretical basis and method for clinical treatment of chest pain.Objective1.In patients with acute myocardial infarction as the research object,using NGF and COX-2 in different TCM syndrome type of the expression is the focus of the study,through the clinical observation method of prospective cohort study,differential expression of NGF,COX-2 in different objective study in patients with acute myocardial infarction among TCM syndromes,to explore the relationship between myocardial sympathetic remodeling and syndrome between,provide an objective basis for TCM diagnosis and treatment and prognosis of acute myocardial infarction.2.Combined with domestic and abroad in recent years for the latest research results of cardiac sympathetic nerve remodeling,by using the model of rats with acute myocardial infarction,detection of acute myocardial infarction rats infarct border zone of myocardial NGF,COX-2 protein level and mRNA expression level of NGF and COX-2 changes in cardiac sympathetic nerve remodeling process and interaction.Method1 Clinical ResearchUsing the method of prospective cohort studies were included in the clinical observation,the diagnosis of acute myocardial infarction,and TCM syndrome with phlegm dampness and blood stasis in patients with type two,the serum NGF in the study,the level of COX-2,a clear expression of the two indexes in patients with acute myocardial infarction,and its relationship between TCM syndrome types.(1)Through non parametric test and other methods,we observed the expression of NGF and COX-2 in serum in acute myocardial infarction patients,and observed their relationship with age,gender,ST segment elevation or non ST segment elevation.(2)Through multivariate logistic regression analysis of NGF and COX-2 levels in patients with different TCM Syndromes of acute myocardial infarction,the correlation between these two indicators and TCM syndromes and possible influencing factors were analyzed.(3)The Spearman rank correlation analysis was used to study the relationship between the serum levels of NGF and COX-2 and the course of acute myocardial infarction in the patients with acute myocardial infarction.(4)The relationship between the serum markers such as NGF and COX-2 and the syndrome type of TCM was observed by ROC curve method.(5)Spearman rank correlation analysis was used to analyze the correlation between NGF and the Gensini score of coronary artery and the level of LDL-C in the serum.2 Animal ExperimentIn this study,the model animals were used to study the histopathology,NGF and COX-2 molecular protein levels and the expression level of mRNA.The objects were divided into 2 groups,including control group and model group,6 rats in each group(successful modeling),in acute myocardial infarction rats after 1 days,7 days,14 in each group were sacrificed 2 rats,myocardium and serum were measured.The experiment is divided into 2 stages:(1)The first stage(modeling stage):model group by ligating the left anterior descending coronary artery,left ventricular anterior wall myocardial ischemia and hypoxia,the ST-T changes and myocardial enzymes increased to confirm the model was successful,normal feeding;control group without ligation of the left anterior descending artery,the remaining steps are the same with the model group,normal feeding.(2)The second stage(animal experiment stage):after the success of the model,respectively in rats with myocardial infarction(MI)after different time points(1 days,7 days,14 days)of the experimental animal were based on specimens were observed in two groups of rats before and after modeling and change the myocardial histopathological manifestation;by using real-time quantitative PCR(qPCR)and Western Blot were observed in the presence of NGF,COX-2 protein and mRNA levels of model group rats myocardial infarction in the marginal division of the abnormal high expression,and compared with control group the expression level;simultaneous determination of expression of NGF,COX-2 mRNA and protein in different periods of model rats and the relationship between the level,to explore the existence of time correlation.Result1 Clinical Research(1)There was no significant difference in the expression of NGF and COX-2 in the different sex of the patients with acute myocardial infarction.(P was 0.515 and 0.366 respectively).There was no significant difference in the expression of NGF and COX-2 in acute ST segment elevation myocardial infarction and non ST elevation myocardial infarction.(P is 0.932 and 0.334,respectively)(2)NGF and COX-2 expression in patients with acute myocardial infarction related to TCM syndrome type,the expression of both in acute myocardial infarction of phlegm stasis syndrome type was significantly higher than that of blood stasis,the difference was statistically significant(P = 0.044 and P<0.001),using multivariate Logistic regression analysis method in the removal of sex and age of course,after effects,still independently associated with NGF and COX-2,both among TCM syndromes(P for trend=0.030 and P for trend<0.001).(3)Multivariate Logistic regression analysis was used to analyze the correlation between NGF and different course of disease.The course of the disease was selected as the dependent variable,and the NGF and the sex,age and TCM syndrome type were independent variables.The results showed that after excluding gender,age and TCM syndromes,the correlation between course of disease and NGF level was still statistically significant(P for trend=0.0497).(4)According to the ability of different markers for differential diagnosis of TCM syndromes,ROC curve analysis showed that COX-2 had the largest area under the curve,and NGF was 0.5996.(5)For non normal distribution data was performed,using Spearman rank correlation analysis between markers,the correlation between NGF and visible COX-2(r=0.234,P=0.017),the relationship between NGF and Gensini score(r=0.065,P=0.0449),between the existence of the relationship between LDL-C and Gensini(r=0.259,P=0.011)between the integral.(6)There was a significant difference in serum NGF between the group and the normal group(P<0.05),which was lower than or in the normal range.The serum COX-2 level of the enrolled patients was significantly different from that of the normal population(P<0.05),which was higher than the normal range.2 Animal Experiments(1)Histopathological results:1 day,7 days and 14 days after coronary artery ligation,the rats in the model group and the sham operation group were killed and 2 rats were observed.The gross specimens and pathological sections of the rats were observed.①From the general specimen,the heart of the control group was complete and dark red.The rats in the model group showed that the myocardium in the vicinity of the ligation was white to the apical direction,and some of the ulcers were necrotic.②From the pathological sections of HE staining of rat myocardial tissue,the myocardium structure of the control group was normal and the boundary was clear,while the myocardium necrosis and rupture,inflammatory cell infiltration,myocardial cell destruction and necrosis of myocardium tissue partially liquefied in the model group.(2)Molecular biological results①qPCR method was used to detect the expression ofinyocardial infarction in the territory of the edge of the two groups of rats NGF and COX-2 mRNA,the expression level of model rats infarct border zone of myocardial NGF and COX-2 mRNA were significantly higher than those in the control group of rats。②qPCR was used to detect the expression of mRNA in different time periods(1 days,7 days and 14 days after operation)in the model group.The expression of NGF and COX-2 mRNA in the infarcted border area increased with time.③Western Blot method was used to detect the relative gray level of NGF and COX-2 in the infarcted border area of two groups.The expression level of NGF and COX-2 protein in model group was higher than that in control group,.④Detection of myocardial infarction model group rats Western Blot method at different time(after 1 days,7 days,14 days)expression of myocardium infarction in NGF and marginal zone COX-2 protein,myocardial infarction 14 days after the group of COX-2 NGF and protein in myocardial tissue of marginal zone infarction in rats was significantly higher than that of the gray value of 1 and 7 days days of rats.Conclusion1 Clinical Research(1)NGF and COX-2 had no differential significance in patients with acute myocardial infarction of different genders.They had no differential significance for acute ST segment elevation myocardial infarction and non ST segment elevation myocardial infarction.(2)NGF,COX-2 expression level was significantly higher than that of blood stasis type in infarction phlegm stasis type acute myocardial,NGF and COX-2 levels were independently associated with TCM Syndromes of acute myocardial infarction,abnormal inflammatory factor and its possible mechanism of phlegm blocking type patients and oxidative stress response related activity.(3)The level of NGF was positively correlated with the course of disease,Gensini score,duration of acute myocardial infarction is longer,the higher the level of NGF in serum;the degree of coronary artery stenosis is more serious,the higher the level of NGF in serum,suggesting that early intervention and improve the coronary artery stenosis to reduce the level of NGF has guiding significance.Gensini score is positively correlated with LDL-C level.The higher the LDL-C level is,the more severe the coronary artery stenosis is.The high LDL-C level plays an important role in coronary atherosclerosis.(4)In this study,the serum levels of NGF in patients with acute myocardial infarction were lower or in the normal range,and the level of COX-2 in the serum was increased.Drugs such as aspirin,statins and beta blockers may have a certain effect on the results of NGF.(5)Through the research on the relationship between NGF,COX-2,and Gensini score of phlegm resistance,found inflammation,oxidative stress and cardiac sympathetic nerve remodeling and myocardial infarction is closely related to the prognosis,may exist in the anti-inflammatory treatment point of intervention for early intervention,so as to improve the new therapy of sympathetic nerve remodeling after myocardial infarction.(6)The active use of Chinese medicine to seek early intervention of phlegm stasis syndrome in patients with treatment of the expression may be down-regulation of NGF and COX-2,improve the myocardial infarction patients sympathetic nerve remodeling,thereby correcting the acute myocardial infarction patients with malignant arrhythmia,heart failure,sudden death and other adverse outcomes,to prevent the occurrence of myocardial infarction after a vicious incident.2 Animal Experiments(1)After acute myocardial infarction,the expression of NGF and COX-2 in myocardial tissue in the marginal zone of myocardial infarction in rats increased.(2)The increase in expression of NGF and COX-2 is time increasing,which may be related to the severity of continuous myocardial ischemia and necrosis and the severity of inflammation.InnovationThe innovation of this study lies in:1.For the first time,two markers of NGF and COX-2 in the process of cardiac sympathetic nerve reconstruction were analyzed with Gensini integral.2.For the first time,the relationship between the two markers of NGF and COX-2 during the process of cardiac sympathetic nerve reconstruction was studied.3.For the first time,the relationship between the elevation of ST segment and the sympathetic nerve remodeling in patients with acute myocardial infarction was studied.4.For the first time on the NGF and COX-2 expression in different patients with acute myocardial infarction among TCM syndromes,Hope that the starting point by using the NGF and COX-2 of two serum markers as traditional Chinese medicine to improve the prognosis of acute myocardial infarction and to expand the traditional Chinese medicine diagnostic method,improve the TCM xiongbi syndrome differentiation theory,provide clinical data support for traditional Chinese medicine intervention of adverse prognosis in patients with acute myocardial infarction.