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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(P<0.01);与H2O2组相比,补阳还五汤组心肌细胞活力显著提高(P<0.01)。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(P<0.01);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(P<0.01)。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(P<0.01)。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(P<0.01)。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。
Objective To investigate the effects of Buyang Huanwu decoction on myocardial cell apoptosis and the expression of 5 lipoxygenase (5-LOX) induced by oxidative stress in neonatal rats. Methods Primary cultured neonatal rat cardiomyocytes were exposed to hydrogen peroxide (H2O2) for 24 hours and then induced by different doses of Buyang Huanwu Decoction. Cell viability was determined by MTT assay. Cardiomyocyte apoptosis was detected by flow cytometry. The expressions of 5-LOX, Bax and Bcl-2 were detected by Western-blot. Results The MTT results showed that compared with the control group and the Buyang Huanwu Decoction group, the viability of cardiomyocytes in H2O2 group was significantly decreased (P <0.01). Compared with H2O2 group, the activity of myocardial cells in Buyang Huanwu Decoction group was significantly increased (P < 0.01). Flow cytometry analysis showed that the apoptosis rate of cardiomyocytes in H2O2 group was (19.47 ± 1.31)%, which was significantly higher than that in control group (1.36 ± 2.30)%, the difference was statistically significant (P <0.01) The percentages of apoptosis in low, medium and high dose groups were (13.25 ± 2.84)%, (11.50 ± 3.43)% and (9.94 ± 4.01)%, respectively, which were significantly lower than those in H2O2 group (19.47 ± 1.31)% Statistical significance (P <0.01). Western-blot results showed that the expression of Bcl-2 protein in H2O2 group was lower than that in control group and BYHWD group, and the expression of 5-LOX and Bax protein were higher than that in control group and BYHWD group Significance (P <0.01). The expression of Bcl-2 protein in Buyang Huanwu Decoction group was higher than that in H2O2 group, while the expression of 5-LOX and Bax protein was lower than that in H2O2 group (P <0.01). Conclusion Buyang Huanwu decoction can significantly inhibit oxidative stress-induced apoptosis in neonatal rat cardiomyocytes in a dose-dependent manner. The effect of Buyang Huanwu Decoction on inhibiting cardiomyocyte apoptosis may be related to the inhibition of 5-LOX expression.