山奈酚、芹菜素、大豆苷元对H_2O_2诱导的心肌细胞凋亡的影响

Effects of kaempferol,apigenin and daidzein on H_2O_2-induced apoptosis in neonatal rat cardiomyocytes

目的观察不同种类黄酮类化合物山萘酚(KA)、芹菜素(AP)、大豆苷元(DA)对H2O2诱导的心肌细胞凋亡及Bcl-2、Bax、Caspase-3蛋白表达的影响。方法取Wistar大鼠新生乳鼠心肌细胞做原代培养,MTT法检测几种黄酮的细胞毒浓度;H2O2(100μmol/L)诱导心肌细胞凋亡;Tunel法、流式细胞仪检测细胞凋亡率;免疫组化方法测定Bcl-2、Bax、Caspase-3蛋白表达。结果MTT法显示各组药物在5、30μmol/L没有显著的细胞毒作用。KA、AP、DA均显著减小H2O2诱导的细胞凋亡率、Caspase-3蛋白表达,增加Bcl-2/Bax比值(P<0.05),其中KA效果最显著(P<0.01)。结论这些黄酮类化合物有抑制H2O2诱导的心肌细胞凋亡作用,其机制与其抗氧化活性有关,可能是通过调节Bcl-2、Bax、Caspase-3蛋白表达,起到心肌细胞保护作用,其中KA效果最好。

Objective To investigate the effects of kaempferol （KA）, apigenin （AP） and daidzein （DA） on cardiomyocytic apoptosis induced by 1-12 02, and the effect of the expressions of Bcl-2, Bax, caspase-3 proteins. Methods Neonatal cardiomyocytes were isolated from one or two days old Wistar rats, and the cytotoxicities of different flavonoids were determined by MTT assay. Cells were treated with 100μmol/L H2O2 to induce apoptosis. Apoptosis rate was determined by flow cytometry（FCM） and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling（TUNEL）. Immunohistochemistry was used for detection of Bcl-2, Bax and caspase-3 protein. Results It showed that each flavonoid did not have noticeable cytotoxicity in 5, 30μmol/L by MTT assay. Kaempferol, apigenin and daidzein significantly decreased the apoptotic rate and the expression of Caspase-3 and increased the ratio of Bcl-2/Bax （P 〈 0.05） compared to model group. Conclusion The resuits suggest that these flavonoids can inhibit the apoptosis induced by H2O2 in neonatal rat cardiomyoeytes. The protective effects could be related to their antioxidant activity and their ability to regulate the expression of Bcl-2, Bax and caspase-3 protein. Kaempferol shows best efllciency.