【摘要】 目的探究柚皮素对缺氧损伤的大鼠心肌细胞是否有保护作用。方法用大鼠心肌细胞系H9c2细胞制备心肌细胞缺氧损伤模型,并用乳酸脱氢酶(LDH)细胞毒性检测试剂盒进行模型鉴定。实验分为5组:对照组,模型组,以及柚皮素低、中、高剂量组(20、40、80 μmol/L)。用CCK-8检测细胞增殖抑制率,间接免疫荧光法检测细胞肥大状况,流式细胞术检测细胞凋亡水平,蛋白质印迹法检测凋亡相关蛋白Bcl-2、Bcl-2关联X蛋白(Bax)、caspase 3的表达水平,qRT-PCR检测血管内皮细胞生长因子(VEGF)、NK2型同源异形框基因5(Nkx2.5)、α-平滑肌肌动蛋白(α-SMA)的mRNA表达水平。结果模型组细胞培养上清中LDH活性高于对照组(P<0.01),表明缺氧损伤心肌细胞建模成功。柚皮素各剂量组细胞增殖抑制率均低于模型组(P<0.05、P<0.01)。与模型组比较,柚皮素各剂量组H9c2心肌细胞肥大状况有所缓解(P<0.05、P<0.01)。柚皮素中、高剂量组凋亡细胞比例均较模型组降低(P均<0.01)。与模型组比较,柚皮素各剂量组凋亡蛋白caspase 3、Bax表达均降低,凋亡抑制蛋白Bcl-2表达均升高(P<0.05、P<0.01)。与模型组比较,柚皮素高剂量组Nkx2.5 mRNA表达水平升高(P<0.01),柚皮素中、高剂量组VEGF mRNA表达水平均升高(P<0.05、P<0.01),柚皮素各剂量组α-SMA mRNA表达水平均降低(P均<0.01)。结论柚皮素对缺氧损伤的大鼠心肌细胞有保护作用。更多还原

【Abstract】 Objective To investigate the protective effect of naringenin on hypoxia-injured myocardial cells. Methods A hypoxia-injured myocardial cell model was established with rat myocardial cell line H9 c2 cells and identified by lactate dehydrogenase(LDH) cytotoxicity test kit. The experiment was divided into 5 groups: control group, model group, and naringenin low-, medium-and high-dose groups(20, 40 and 80 μmol/L). Cell proliferation inhibition rate was detected by cell counting kit 8; cell hypertrophy was detected by indirect immunofluorescence; cell apoptosis was detected by flow cytometry; the protein expression of apoptosis-related proteins B-cell lymphoma 2(Bcl-2), Bcl-2 associated X protein(Bax) and caspase 3 was detected by Western blotting; and the m RNA expression of vascular endothelial growth factor(VEGF), NK2 homeobox 5(Nkx2.5) and alpha-smooth muscle actin(α-SMA) in H9 c2 cells was detected by q RT-PCR. Results Compared with the control group, the activity of LDH in cell supernatant of the model group was significantly increased(P<0.01), indicating that the hypoxia-injured myocardial cell model was successfully constructed. Compared with the model group, the inhibition rate of cell proliferation was significantly decreased in each dose group of naringenin(P<0.05, P<0.01); the hypertrophy of H9 c2 cells in each dose group of naringenin was significantly alleviated(P<0.05, P<0.01); and the proportion of apoptotic cells was significantly decreased in the medium-and high-dose naringenin groups(both P<0.01). Compared with the model group, the expression levels of apoptotic proteins caspase 3 and Bax in each dose group of naringenin were significantly decreased, while the expression levels of apoptosis inhibition-related protein Bcl-2 were significantly increased(P<0.05, P<0.01). Compared with the model group, the m RNA expression level of Nkx2.5 in high-dose naringenin group was significantly increased(P<0.01), the m RNA expression levels of VEGF were significantly increased in medium-and high-dose naringenin groups(P<0.05, P<0.01), and the m RNA expression levels of α-SMA were significantly decreased in each dose group of naringenin(all P<0.01). Conclusion Naringenin has a protective effect on hypoxia-injured myocardial cells of rats.更多还原