心脉通抗氧化应激所致心肌细胞凋亡的机制探讨

Anti-apoptosis Mechanism of Serum Containing Xin Mai Tong Decoction in Myocardial Cell Under Oxidative Stress

目的探讨心脉通含药血清抗氧化应激所致心肌细胞凋亡的分子机制。方法培养乳鼠心肌细胞,采用400μmol/LH2O2建立氧化应激损伤导致心肌细胞凋亡模型。采用MTT法检测细胞活力改变,TUNEL法检测心肌细胞凋亡,免疫细胞化学方法检测HSP70、NF-κB、caspase-3在细胞内表达。结果H2O2作用1h对心肌无损伤作用,HSP70表达轻度增加。H2O2作用6h可导致心肌细胞凋亡,心脉通含药血清可减轻H2O2导致的细胞活力下降,显著抑制心肌细胞凋亡,明显增加HSP70在细胞内的表达,减少NF-κB核转位,减少心肌细胞内caspase-3表达。结论心脉通含药血清对H2O2导致的心肌细胞凋亡具有明显的抑制作用,其可能机制是增加HSP70蛋白表达,抑制NF-κB活化,减少心肌细胞内caspase-3表达。

Objective To explore the anti - apoptosis mechanism of serum containing Xin Mai Tong decoction in myocardial cell under oxidative stress. Methods The neonate rat myocardial cell was cultivated and the model of myocardial cell apoptosis was induced with 400 μmol/L H2O2. The cell viability was determined by MTT method, myocardial cell apoptosis was determined by TUNEL method, and the intra - cellular expression of HSP70, NF - κB and caspase - 3 in myocardial cell was determined by immunocytochemistry. Results Administration of 400 μmol/L H2O2 for 1 hour was harmless to myocardial cell, the expression of HSP70 being slightly increased. However, apoptosis was induced in myocardial cells at the 6 th hour after 400μmol/L H2O2 administration. The serum containing Xin Mai Tong significantly attenuated the decrease of cell viability and apoptosis which was induced by oxidative stress. Increased expression of HSP70 was induced by the serum containing Xin Mai Tong, and the activation of NF - κB and caspase - 3 were significantly inhibited. Conclution The serum containing Xin Mai Tong decoction shows a significant inhibitory effect on apoptosis induced by H2O2, and the mechanism may be through increasing protein creasing the expression of caspase - 3. expression of HSP70, inhibiting the activation of NF - κB and de-