当归红芪超滤膜提取物抑制过氧化氢诱导的大鼠心肌细胞凋亡及相关基因表达

The Ultra-Filtration Extract From the Mixture of Angelica Sinensis and Hedysarum Polybotrys Can Inhibit Cardiomyocyte Apoptosis Induced by Hydroperoxide and Related Gene Expression in Rats

目的探讨当归红芪超滤膜提取物对心肌细胞凋亡相关基因bc l-2、bax蛋白表达的影响。方法用高浓度的过氧化氢(400μmol/L)在W istar乳鼠原代培养的心肌细胞上建立细胞凋亡模型,用不同浓度的当归红芪超滤物(3.75、7.5、15 g/L)进行治疗。光镜下观察细胞形态;逆转录-多聚酶链反应和蛋白印迹法检测bc l-2、bax蛋白在心肌细胞中的表达情况。结果与正常对照组相比,高浓度过氧化氢损伤组bc l-2 mRNA及蛋白表达显著降低(P<0.05);bax mRNA及蛋白表达显著增加(P<0.05);bc l-2/bax比值显著降低(P<0.01),差异具有统计学意义。与损伤组比较,高浓度和中浓度药物治疗组bc l-2 mRNA及其蛋白表达显著增加(P<0.05);bax mRNA及其蛋白表达显著降低(P<0.05);bc l-2/bax比值显著增加(P<0.01),差异具有统计学意义。低浓度药物治疗组则对bc l-2、bax表达无显著改变。结论当归红芪超滤膜提取物可通过上调bc l-2/bax的比值抑制心肌细胞的凋亡,对心肌细胞具有抗凋亡的保护作用。

Aim To study the effect of ultra-filtration extract from the mixture of angelica sinensis and hedysarum polybotrys on the expressions of protein and mRNA of bcl-2 and bax gene in the apoptotic myocardial cells. Methods H2O2 of 400μmol/L was used to build an oxidative stress-induced injury model in neonatal rat cardiomyocytes and then cardiomyocytes of each treatment group were treated with the Ultra-filtration extract （ 3.75,7.5, 15 g/L）. Morphological changes of cardiomyocytes were observed in microscope. The levels of bcl-2 and bax expression in cardiomyocytes were measured by RT-PCR and Western-blot. Results In cardiomyocytes of model group, the expression of bcl-2 and the rate of bcl-2/bax were decreased, and the expression of bax was increased significantly, compared with normal group （ P 〈 0.05 ）. In cardiomyocytes of the high dosage group and the middle dosage group of the treatment, the bcl-2 expression was increased （ P 〈 0.05 ） , the bax expression was decreased （ P 〈 0.05 ）, and the rate of bcl-2/bax was increased （ P 〈 0.05）, compared with model group. In cardiomyocytes of the little dosage group of the treatment, the expressions of bcl- 2 and bax and the rate of bcl-2/bax were not changed significantly. Conclusion Ultra-filtration extract from the mixture of angelica sinensis and hedysarum polybotrys can effectively prevent and treat myocardial cells apoptosis, and its mechanism may be associated with up-regulation of the rate of bcl-2/bax .