银杏黄酮对缺氧复氧心肌H9c2细胞损伤的保护作用及其机制

Protective effect and mechanism of ginkgo biloba on hypoxia reoxygenation-induced myocardial H9c2 cells damage

目的探讨银杏黄酮对缺氧复氧损伤大鼠心肌H9c2细胞的保护作用,并探讨其机制。方法培养H9c2细胞,造成缺氧复氧损伤模型,分为对照组、实验组、银杏黄酮低浓度组及银杏黄酮高浓度组,CCK8法观察细胞活性,测各组细胞上清液乳酸脱氢酶(LDH)及细胞超氧化物歧化酶(SOD)、丙二醛(MDA)变化,Hoechst染色检测凋亡细胞比例,电镜观察细胞超微结构改变,Western blot分析Bcl-2及Bax的变化。结果CCK8结果显示,用1、10μg/mL浓度的银杏黄酮处理后,H9c2细胞存活率分别提高到(54.80±4.27)%和(68.04±6.85)%,与实验组(31.37±3.51)%比较差异具有统计学意义(P<0.05);用1、10μg/mL浓度的银杏黄酮处理后,H9c2细胞存活率分别提高到(54.80±4.27)%和(68.04±6.85)%,与实验组31.37%±3.51%比较差异具有统计学意义(P<0.05);Hoechst染色结果显示,实验组细胞凋亡率为(69.09±5.31)%,与对照组(4.57±2.34)%比较差异具有统计学意义(P<0.05),银杏黄酮低浓度组及银杏黄酮高浓度组细胞凋亡率分别为(45.88±5.53)%及(23.63±4.27)%,与实验组比较差异均有统计学意义(P<0.05)。此外,银杏黄酮改善了细胞形态,Western Blot检测结果显示,实验组中Bcl-2蛋白相对表达量为0.56±0.35,与对照组(相对表达量为1.63±0.54)相比差异有统计学意义(P<0.05);低、高浓度银杏黄酮组Bcl-2蛋白相对表达量分别为0.80±0.16、1.22±0.18,与实验组比较差异均有统计学意义(P<0.05);实验组中Bax蛋白相对表达量为0.89±0.11,与对照组(相对表达量为0.21±0.06)相比差异有统计学意义(P<0.05),低、高剂量银杏黄酮组Bax蛋白的相对表达量(0.52±0.10、0.30±0.12)与实验组相比差异有统计学意义(P<0.05)。结论银杏黄酮可以抑制缺氧复氧诱导的H9c2细胞氧化损伤及凋亡的发生,在预防和治疗心肌缺血再灌注损伤疾病方面具有一定的应用前景。

Objective To investigate the protective effect and mechanism of ginkgo biloba on hypoxia reoxygenation-induced myocardial H9c2 cells damage and explore its mechanisms.Methods Myocardial H9c2 cells were cultured and hypoxia reoxygenation models were constructed.Cells were devided into control group,experimental group,low concentration and high concentration of ginkgo biloba groups.Cell survival rate was detected by CCK8 colorimetry.The changes of lactate dehydrogenase(LDH)in cell supernatant,superoxide dismutase(SOD)and malondialdehyde(MDA)in cells were measured.Apoptotic cells proportion were detected by Hoechst staining.Cell ultrastructure changes were observed by electron microscope.The expression of Bcl-2 and Bax were tested by western blot assau.Results The CCK-8 results showed that cortical neurons survival rate increased to 54.80%±4.27%and 68.04%±6.85%after 1μg/ml and 10μg/ml ginkgo biloba treatment.Hoechst staining results showed that cells apoptosis rate of experimental group was[(69.09±5.31)%],compared with the control group[(4.57±2.34)%].Cells apoptosis rate in low concentration and high concentration of ginkgo biloba groups was(45.88±5.53)%and(23.63±4.27)%,respectively,compared with the experimental group.In addition,ginkgo biloba improved the cell morphology.Western blot assay showed that the expression of Bcl-2 protein in the experimental group was lower than that in control group.The expression of Bcl-2 in low concentration and high concentration of ginkgo biloba groups was higher than that in experimental group.The expression of Bax protein in the experimental group was increased compared with the control group.The expression of Bax in low concentration and high concentration of ginkgo biloba group was reduced compared with the experimental group.Conclusion Ginkgo biloba could inhibit hypoxia reoxygenation-induced oxidative damage and apoptosis in myocardial H9c2 cells.