The Effects Of SiMiaoYongAn Decoction With ShengMai Decoction On Anoxic Cultured Neonatal Rat Myocardial Apoptosis

Objective:To investigate the effects of SiMiaoYongAn decoction withShengMai decoction on apoptosis of Cardiac Myocytes injured afterHypoxia/Reoxygenation.Methods：In this experiment，SiMiaoYongAn decoction with ShengMaidecoction was decocted and made by alcohol sedimentation,the combinationof trypsin and collagenase was to isolate neonatal rat myocardial cells, whichwere purified twice by differential adhesion method and identified byImmunocytochemical methods,and these cells were observed under theinverted phase contrast microscope during different time,then taken picturesand videos. The myocardial hypoxia/reoxygenation model was establishedwith high purity nitrogen, cultured neonatal rat myocardial cells weresubjected to hypoxia4h and reoxygenation2h.Primary cultured neonatal ratcardiomyocytes were randomly divided into three groups,control group:themyocardial cells were normally cultured; model group: they were injured byhypoxia/reoxygenation, but without the alcohol sedimentation solution ofSiMiaoYongAn decoction with ShengMai decoction;treatment group:theywere interfered by hypoxia/reoxygenation and then dealed with thesolution.The myocardium viability was detected by MTT colorimetricmethod,and aspartate aminotransferase(AST) was determined by automaticbiochemial analyzer,and the rate of myocardium was measured with flowcytometry.Results:(1)Under the inverted phase contrast microscope, primarycultured neonatal rat myocardial cells stretched the parapodium out and hadspontaneous beatings, as well as these cells were identified into myocardialcells by immunocytochemistry.(2) The effects of SiMiaoYongAn decoctionwith ShengMai decoction on anoxic cultured neonatal rat myocardial apoptosis:①The myocardium viability detected by MTT colorimetricmethod:compared with control group, the OD of model group and treatmentgroup was significantly lower but that of treatment group was higher thanmodel group，which showed live Cardiac Myocytes of the model andtreatment groups decreased while those of treatment group were more thanmodel group.By the single factor analysis of variance,model group andtreatment group had a significant difference (P <0.05) compared with normalcontrol group,which was statistical significance; The differences of modelgroup and treatment group was significant (P <0.05), there was also statisticalsignificance;②the content of aspartate aminotransferase(AST) in culturemedia determined by automatic biochemial analyzer:myocardial culture mediaof control group,model group,and treatment group for redundancy afterhypoxia/reoxygenation treatment,the AST content of control group was (1.02+0.45),the level of model group was (3.96+0.88), and in the treatment groupit was (2.55+0.67), control group was lower than the other tow, but modelgroup was higher compared with treatment group. The result showed that thecontent of AST had a significant difference (P <0.05).③the rate ofmyocardial apoptosis measured with flow cytometry:The myocardial cellscultured in the normal condition display apoptosis,the apoptosis rate was(13.02+0.65)%; and they were respectively (28.56±0.92)%and (16.74±0.83)%between model group and treatment group.The results showed therewere significant difference among three different groups (P<0.05).Consclution: the alcohol sedimentation solution of SiMiaoYongAndecoction with ShengMai decoction played a protective role on myocardialapoptosis,which may be relative with the improvement of the myocardiumviability and the decline of the AST content in culture media induced byhypoxia/reoxygenation treatment;the protection of the alcohol sedimentationsolution on myocardial apoptosis provides a new way for the treatment ofcoronary heart disease and heart failure. But the specific mechanism of theprotection is yet to be further experiments, and we need to research concrete and effective material composition thoroughly.