补阳还五汤对氧化应激诱导心肌细胞凋亡及5-LOX表达的影响

目的探讨补阳还五汤对氧化应激诱导乳鼠心肌细胞凋亡及5脂氧合酶(5-LOX)表达的影响。方法 SD乳鼠心肌细胞原代培养,不同剂量补阳还五汤预处理24 h后过氧化氢(H2O2)诱导心肌细胞氧化损伤。采用MTT法测定细胞活力,流式细胞术检测心肌细胞凋亡情况,Western-blot检测5脂氧合酶(5-LOX)、Bax和Bcl-2的表达。结果 MTT结果显示,与对照组及补阳还五汤组比较,H2O2组心肌细胞活力显著降低(P0.01);与H2O2组相比,补阳还五汤组心肌细胞活力显著提高(P0.01)。流式细胞仪分析显示,H2O2组心肌细胞凋亡率为(19.47±1.31)%,明显高于对照组的(1.36±2.30)%,差异有统计学意义(P0.01);补阳还五汤低、中、高剂量组的细胞凋亡百分率分别为(13.25±2.84)%、(11.50±3.43)%、(9.94±4.01)%,明显低于H2O2组(19.47±1.31)%,差异有统计学意义(P0.01)。Western-blot结果显示,H2O2组心肌细胞Bcl-2蛋白表达比对照组及补阳还五汤组减少,5-LOX及Bax蛋白表达比对照组及补阳还五汤组高,差异均有统计学意义(P0.01)。不同剂量的补阳还五汤组Bcl-2蛋白表达高于H2O2组,5-LOX及Bax蛋白表达比H2O2组少(P0.01)。结论补阳还五汤能显著抑制氧化应激诱导的乳鼠心肌细胞凋亡,并且呈剂量依赖性,补阳还五汤抑制心肌细胞凋亡的作用可能与抑制5-LOX表达有关。

Effect of Buyang huanwu decoction on oxidative stress-induced neonatal rat cardiomyocytes apoptosis and expression of 5-LOX

Objective To investigate the effect of Buyang huanwu decoction（BYHWD） on oxidative stress-induced neonatal rat cardiomyocytes apoptosis and expression of 5-lipoxygenase（5-LOX）. Methods After pretreated with BYHWD or not pretreated with BYHWD for 24 hours, H2O2was used to induce oxidative stress injury in primary cultured cardiomyocytes.Cell viability was detected by MTT assay.Cardiomyocyte apoptosis was determined by flow cytometry.Expression of 5-LOX, Bax and Bcl-2 were detected by Western-blot. Results MTT assay showed that in H2O2group, cell viability was significantly decreased compared with control group and BYHWD group（P 0.05）.As compared with H2O2group,cell viability improved significantly in BYHWD group（P〈0.05）.Flow cytometry analysis showed that the percentage of apoptosis cells in H2O2group was（19.47±1.31）%, which was significantly higher than control group（1.36±2.30）%（P〈0.01）.The percentage of apoptosis cells in BYHWD low-dose group,middle-dose group and high-dose group were（13.25±2.84）%,（11.50±3.43）% and（9.94±4.01）%, respectively, which were significantly lower than H2O2group（P〈0.01）.Western-blot showed that the Bcl-2 expression level of H2O2group was significantly lower than that of control group and BYHWD group,and the 5-LOX and Bax expression level was significantly higher than those of control group and BYHWD group（P〈0.05）. The Bcl-2expression level of BYHWD group was obviously higher than H2O2 group, and the 5-Lox and Bax expression levels were obviously lower than H2O2group（P〈0.05）. Conclusion BYHWD can inhibit oxidative stress induced neonatal rat cardiomyocytes apoptosis with a dosedepending manner. This Pharmacological effect of BYHWD may via the suppression of 5-LOX expression.