7-羟乙基白杨素对心肌细胞缺氧损伤的保护作用及机制研究

Protective effect and mechanism of 7-hydroxyethyl chrysin on hypoxia injury of cardiomyocytes

目的探究7-羟乙基白杨素对心肌细胞缺氧损伤的保护作用及其信号转导机制。方法将心肌细胞株H_(9)C_(2)传代后随机分为正常对照组(常氧条件培养24 h)、模型组(1%O_(2)浓度缺氧培养24 h)、白杨素组(1%O_(2)浓度缺氧培养24 h,1.0×10^(-6)mol·L^(-1)白杨素处理)、7-羟乙基白杨素组(1%O_(2)浓度缺氧培养24 h,1.0×10^(-6)mol·L^(-1)7-羟乙基白杨素处理)。用细胞计数试剂盒-8(CCK-8)法检测细胞增殖情况;用WST-1法测定超氧化物歧化酶(SOD)含量;用钼酸铵法测定过氧化氢酶(CAT)含量;用蛋白质印迹法检测血红素加氧酶1(HO-1)、过氧化氢酶(CAT)蛋白表达水平。结果正常对照组、模型组、白杨素组和7-羟乙基白杨素组的细胞存活率分别为(100.00±4.48)%、(61.95±2.14)%、(86.01±3.60)%和(93.16±2.91)%;SOD活力分别为111.43±3.06、196.12±12.09、162.48±7.64和141.55±2.82;CAT活力分别为3.40±0.32、9.91±0.39、5.51±0.20和4.41±0.43;HO-1蛋白表达水平分别为0.58±0.01、1.22±0.02、0.68±0.01和0.34±0.01;SOD蛋白表达水平分别为0.38±0.01、1.07±0.01、0.92±0.01和0.49±0.01;CAT蛋白表达水平分别为0.73±0.02、1.10±0.02、0.94±0.02和0.67±0.01。模型组的上述指标与正常对照组比较,7-羟乙基白杨素组、白杨素组上述指标与模型组比较,7-羟乙基白杨素组上述指标与白杨素组比较,差异均有统计学意义(均P<0.01)。结论7-羟乙基白杨素通过延缓核因子-E2相关因子/抗氧化应答元件信号途径的激活而提高了心肌细胞的抗氧化损伤能力。

Objective To investigate the anti-hypoxia injury effects of 7-hydroxyethyl chrysin(7-HChr)on cardiomyocytes H_(9)C_(2)and clarify its molecular mechanism.Methods H_(9)C_(2)cells were subcultured and randomly divided into control group(cultured under normoxia for 24 hours),model group(cultured under 1%O_(2)for 24 hours),chrysin group(1%O_(2)for 24 hours,1.0×10^(-6)mol·L^(-1)chrysin),and 7-hydroxyethyl chrysin group(1%O2 for 24 hours,1.0×10^(-6)mol·L^(-1)7-hydroxyethyl chrysin).Cell counting kit-8(cck-8)test was used to detect cell proliferation;WST-1 was used to determine the level of superoxide dismutase(SOD);ammonium molybdate method was used to detect the activity of catalase(CAT);Western blot method was used to detect heme oxygenase 1(HO-1)and CAT protein expression.Results The cell survival rates of control group,model group,chrysin group and 7-hydroxyethyl chrysin group were(100.00±4.48)%,(61.95±2.14)%,(86.01±3.60)%and(93.16±2.91)%,respectively;the SOD activity were 111.43±3.06,196.12±12.09,162.48±7.64 and 141.55±2.82,respectively;the CAT activity were 3.40±0.32,9.91±0.39,5.51±0.20 and 4.41±0.43,respectively;the HO-1 protein expressions were 0.58±0.01,1.22±0.02,0.68±0.01 and 0.34±0.01,respectively;the SOD protein expressions were 0.38±0.01,1.07±0.01,0.92±0.01 and 0.49±0.01,respectively;the CAT protein expressions were 0.73±0.02,1.10±0.02,0.94±0.02 and 0.67±0.01,respectively.Comparison between the model group and control group,7-hydroxyethyl chrysin group and chrysin group,and comparison between the 7-hydroxyethyl chrysin group and chrysin group indicated that the differences of above indicators were significant(all P<0.01).Conclusion 7-hydroxyethyl chrysin enhances the ability of cardiomyocytes H9C2 to resist oxidative damage by delaying the activation of nuclear factor-E2-related factor/antioxidant response element signaling pathway.