| PurposeCardiomyocyte apoptosis in certain physiological and pathological conditions, the signal transduction pathway, activation of cell "suicide" program, a series of genes under the control of programmed cell death. Recent studies show that apoptosis in myocardial cells play an important role, in the heart of the physiological and pathological development process and to maintain normal cardiac morphology, is considered the heart by compensatory changes in the pathological changes in the development of the cytological basis." The prevalence of cardiac myocyte apoptosis in myocardial infarction, myocardial ischemia-reperfusion injury, heart failure, dilated cardiomyopathy, and adriamycin-induced cardiomyopathy and other cardiovascular diseases, ischemic myocardial infarction myocardial cell death one of the ways. In recent years, the study found that cardiac myocyte apoptosis, inflammatory cytokines involved in a series of pathological changes in left ventricular remodeling after myocardial infarction, heart failure, cardiac myocyte apoptosis is to expand the myocardial infarct size is an important factor not only affects myocardial infarct size, and contribute to myocardial remodeling, myocardial infarction, early and late apoptotic phenomenon exists.Apoptosis by a variety of gene regulation, this process is affected by a variety of pro-apoptotic and anti-apoptotic protein regulation, including pro-apoptotic gene Bax and inhibition of apoptosis gene Bcl-2 is considered to be closely related with cell apoptosis, Experimental results show that the Bcl-2 and Bax gene involved in the regulation of cardiac hypertrophy in cardiomyocyte apoptosis. Bcl-2 which inhibit a variety of factors such as oxygen free radicals and p53-induced apoptosis. Bax pro-apoptotic factor, and its overexpression will accelerate -3 mediated by interleukin-induced apoptosis. When the trigger factors exist, of Bcl-2/Bax ratio determines the cells is survival or death.The p53 gene is an important apoptosis-related genes, divided into two types: wild-type and mutant. Mutant p53 can promote cell growth, and participation in various tumors. The main function of the wild type p53 gene is involved in the negative regulation of cell growth and limit cell growth and division. Recent studies have found that the p53 gene not only with the occurrence of many tumors, related to the development and is involved in the occurrence of apoptosis in the cardiovascular system. Research has shown that expression of the P53 gene in acute ischemia and ischemia -reperfusion injury in cardiac myocyte apoptosis plays an important role.One of the p38 activation of the mitogen-activated protein kinase (the mitogen activated protein kinase MAPKs) subfamily members, the full name of p38MAPK. Studies suggest that p38 is a common pathway of cell proliferation and differentiation, apoptosis and necrosis signaling, with the induction of apoptosis effect. p38 is the most extensive study of cardiac myocyte apoptosis, which include p38α, p38β, p38γ, p38δ subtypes. The study of p38MAPK and the beginning of cardiac myocyte apoptosis is closely related to its activation may be in the process of cardiac myocyte apoptosis plays a key role.Acetaldehyde reductase 2 (ALDH2) is a acetaldehyde oxidation enzymes in mitochondria, and it is closely related to biological oxidation, prevents acetaldehyde on membrane lipid peroxidation, reducing the metabolism of reactive oxygen species products cell damage. ALDH2 is widely distributed in the human liver, kidney, heart, lung, brain and other organizations, is one of the main enzyme aldehyde oxidation. Experiments show that ALDH2 caused by oxidation to stimulate nerve cells, endothelial cell injury has a protective effect in recent years also found that ALDH2 participate in the cardioprotective effect was found to activate ALDH2 can reduce cardiac ischemic injury, thus ALDH2 as cardiovascular and cerebrovascular diseases biochemical markers of attention. The foreign scholar studies have shown that high expression of ALDH2 also slow the process of apoptosis.Buyanghuanwu Decoction "Correction of Medicine" from the Qing Dynasty Wang Qing-ren is Yiqihuoxue represented party, and its reuse Astragalus King, make up their strength so that the gas Wang blood line, flow body; Chuanxiong, red peony root, normalized tail Tameomi, nourishing and Blood Circulation; peach kernel, safflower for the rank and file, breaking the knot stasis; earthworm to make Tom Lee meridian, is commonly used prescription of Chinese medicine clinical treatment of cardiovascular and cerebrovascular diseases. Bu Yang Huan Wu Tang modern experimental studies have shown that:Buyanghuanwu Decoction has a therapeutic effect on a variety of cardiovascular and cerebrovascular diseases. Bu Yang Huan Wu Tang can inhibit brain ischemia-reperfusion injury; protect the artery atherosclerosis sclerosis mouse model of endothelial function, inhibit the development of atherosclerosis; to improve chronic heart failure patients with left ventricular systolic and diastolic function; inhibition of hypoxia reoxygenation-induced cardiomyocyte apoptosis. Our previous studies showed that the Bu Yang Huan Wu Tang can significantly improve left ventricular structure and function, inhibition of interstitial collagen deposition in the inhibition of cardiomyocyte apoptosis, reduction of myocardial fibrosis. Cardiomyocyte apoptosis plays an important role in the development of ventricular remodeling after myocardial infarction Buyanghuanwu decoction anti-myocardial infarction left ventricular remodeling, inhibition of the mechanism of cardiac myocyte apoptosis deserve further study.ObjectivePrevious studies show that Buyanghuanwu decoction for rat myocardial infarction ventricular remodeling. This study by observing Yang Huan Wu Decoction on myocardial apoptosis after myocardial infarction, as well as of ALDH2, Bax and Bcl-2, p53, expression of p38 protein, designed to reduce cardiac myocyte apoptosis explore BHD Wu Tang is to improve the mechanisms of ventricular remodeling after myocardial infarction. Its TCM clinical treatment of cardiovascular diseases, especially after the Chinese anti-myocardial infarction ventricular remodeling to provide further experimental evidence.MethodThe use of SPF male Wistar rats, ligation of the left main coronary artery causing myocardial infarction model. Myocardial infarction model rats were randomly divided into three groups:model control group, the perindopril group, Bu Yang Huan Wu Tang group; with sham group, a total of four groups (n= 12),48. Intragastric administration of Bu Yang Huan Wu Tang group:5g/kg · d TCM decoction of gavage; in the perindopril group:1g/L perindopril suspension 10mg/kg d dose; sham operation group and model control group:normal drinking water for consecutive 12 weeks. Figure:The left ventricular long axis view of the probe frequency is 12 MHz, using a standard apical four-chamber view, five chamber view surface 12 weeks after the detection of Doppler ultrasound echocardiography, while recording the ECG R wave peak end-diastolic careful observation of the morphology of the heart to the end of the T wave, systolic amplitude of wall motion and systolic thickening. Morphological indicators include:left ventricular end-diastolic diameter (LVDd), left ventricular end systolic diameter (LVDs), and then calculate the value of the left ventricular ejection fraction (LVEF) values and left ventricular fractional contraction (LVFS) were compared among groups rat heart morphology and cardiac function. TUNEL staining to detect myocardial apoptosis; immunohistochemical method for the determination of the myocardial tissue of Bax, Bcl-2 protein content; Western-Blot determination of myocardial tissue of p53, p38, ALDH2 protein content. The data were analyzed using SPSS 13.0 statistical software, the result is expressed as:mean ± standard deviation (± S), the results were compared using one-way ANOVA. Homogeneity of variance, the two groups were compared using LSD, when heterogeneity of variance, using the Welch robust estimation, and then using Dunnett’sT3 or Tamhane’s T2 method pairwise comparisons., P<0.05, said that there are statistically significant. Result 1 Results of cardiac ultrasonographyVarious indicators of group differences in LVDd (F= 121.367, P＜0.001), LVDs (F= 75.411, P<0.001), EF (F=77.340, P<0.001), FS (F=135.152, P<0.001) andcompared to the sham operation group, model control group, Yang Huan Wu Tang group and perindopril group, LVDd and LVDs increased significantly (P= 0.000), model control group increased the most significant; model control group, LVDd and LVDs were significantly greater thanBu Yang Huan Wu Tang group and perindopril group (P<0.001), Bu Yang Huan Wu Tang group and perindopril group differences. Compared with the sham group Bu Yang Huan Wu Tang group and perindopril group EF and FS values difference not statistically significant (P< 0.001), EF and FS values of the model control group was significantly less than BHD Huan Wu Tang group and perindopril group:EF (P=0.016, P=0.007), FS (P=0.003, P=0.002).2 TUNEL assay results of myocardial apoptosisSham operation group, no significant apoptosis in myocardial cells, the microscope field of vision was dark green; model control group found a large number of apoptotic myocardial cells, apoptotic cells were bright green, apoptotic cells were located in the edge of the infarcted area; Buyanghuanwu decoction groupperindopril group shows a small amount of apoptosis in cardiac myocytes, the degree of the control group compared with the model significantly reduced (P<0.001), but it does not completely prevent its occurrence.3 Results of immunohistochemical method for the determination of the myocardial tissue of Bax, Bcl-2 protein contentOnly a small amount of Bax and Bcl-2 expression in the myocardial tissue of sham operation group; model control group of Bax, Bcl-2 protein expression more, the ratio of Bax/Bcl-2 ratio was significantly greater than the sham operation group rats (P< 0.001); Bu Yang Huan Wu Tang group and perindopril group with the sham group compared to the Bax/Bcl-2 also increased (P= 0.005, P= 0.001), MI of Bax and Bcl-2 protein expression were increased, but even more significant increase in Bax protein, Bcl-2 protein expression may increase and myocardial cells stimulated by the ischemic process and increased expression of reactive, in order to improve the intrinsic resistance of the cells is related to;The expression was mainly located in the marginal zone of myocardial infarction. And model control group, Yang Huan Wu Tang group and perindopril intervention downregulation of Bax protein expression (P < 0.001, P= 0.002), and mild increase trend of Bcl-2 protein expression (P= 0.002, P= 0.013), reducing the ratio of Bax/Bcl-2 (P= 0.002, P= 0.013), thereby reducing cardiac myocyte apoptosis.4 Result of Western-Blot determination of p53 protein content in myocardial tissueModel control group, p53 protein expression was significantly increased, the gray value is significantly higher than sham operation group (P=0.001); Buyanghuanwu decoction of the gray value of the perindopril group compared with the sham group,but significantly lower than control group (P=0.001).5 Result of Western-Blot determination of p38 protein content in myocardial tissueModel control group, Yang Huan Wu Tang group, perindopril group, p38 protein expression was significantly higher than the sham group, striped gray values are statistically different (P< 0.001), in which sham operation groupthe gray value is obviously higher than that of Buyanghuanwu decoction group and perindopril group (P<0.001,P=0.001).6 Result of Western-Blot determination of ALDH2 protein content in myocardial tissueModel control group, only a trace ALDH2 expression, striped gray level was significantly lower than the sham operation group (P< 0.001), Buyanghuanwu decoction group and perindopril group ALDH2 expression levels compared with the model control group compared with the sham group (P<0.001,P=0.001). The results indicate that no drug interventions after myocardial infarction, myocardial cells ALDH2 downregulation of Bu Yang Huan Wu Tang can mcrease myocardial infarction after myocardial cells the expression of ALDH2, thereby reducing cardiac myocyte apoptosis.ConclusionBuyanghuanwu decoction rate of reduction of Bax, p53, p38 MAPK expression and up-regulated the expression of Bcl-2, ALDH2 expression of cardiac myocytes after myocardial infarction pro-apoptotic and anti-apoptotic protein expression have been regulated, thereby reducing the myocardial cellsapoptosis, anti-ventricular remodeling. |
