茶多酚对H9C2大鼠心肌细胞缺氧/复氧损伤的保护作用机制

Study on the protective effect and mechanism of tea polyphenols on H9C2 rat cardiomyocytes injury induced by hypoxia/reoxygenation

目的探究茶多酚对H9C2大鼠心肌细胞缺氧/复氧损伤的保护作用及机制。方法实验开展时间为2017年12月至2019年3月,分为对照组、缺氧/复氧组、低浓度茶多酚组和高浓度茶多酚组,对照组予以常规孵箱培养H9C2大鼠心肌细胞,缺氧/复氧组利用缺氧孵箱和常规孵箱交替模拟缺血再灌注环境培养H9C2大鼠心肌细胞,低浓度茶多酚组在缺氧孵箱培养后在常规孵箱培养前应用2 mg/mL的茶多酚,高浓度茶多酚组加入4 mg/mL的茶多酚。利用CCK-8法检测48 h培养后各组细胞活性;利用试剂盒检测48 h培养后各组丙二醛(MDA)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和谷胱甘肽过氧化物酶(GSH-Px)等氧化还原指标和髓过氧化物酶(MPO)、白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)和IL-6等炎症指标;利用免疫印迹技术检测丝裂原活化蛋白激酶(MAPK)和核因子κB通路相关分子。结果与缺氧/复氧组相比,茶多酚组细胞活性增强(P<0.05);与缺氧/复氧组相比,茶多酚组MDA含量减少(P<0.05),SOD、CAT和GSH-Px含量增加(P<0.05);与缺氧/复氧组相比,茶多酚组MPO、IL-1β、TNF-α和IL-6含量减少(P<0.05);与缺氧/复氧组相比,茶多酚组MAPK和核因子κB通路相关分子表达被抑制(P<0.05)。结论茶多酚能够通过丝裂原活化蛋白激酶和核因子κB通路对H9C2大鼠心肌细胞缺氧/复氧损伤发挥保护作用。

Objective To examine the protective effect and mechanism of tea polyphenols on H9C2 rat cardiomyocytes injury induced by hypoxia/reoxygenation.Methods The experiments were conducted from December 2017 to March 2019.H9C2 rat cardiomyocytes were randomly divided into control group,hypoxia/reoxygenation group,low concentrations of tea polyphenols group and high concentrations of tea polyphenols group.H9C2 rat cardiomyocytes in the control group were cultured in conventional incubator,and cells in the hypoxia/reoxygenation group were cultured in hypoxia incubator and conventional incubator alternatively to simulate ischemia reperfusion injury of skeletal muscle cells.Cells in the low and high concentrations of tea polyphenols groups were treated with 2 mg/mL and 4 mg/mL tea polyphenols after incubation in hypoxic incubator and before incubation in conventional incubator.CCK-8 assay was used to test cell viability.Indicated test kits were used to examine the content of redox indexes such as malondialdehyde(MDA),superoxide dismutase(SOD),catalase(CAT)and glutathione peroxidase(GSH-Px)and inflammatory indexes such as myeloperoxidase(MPO),interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6).Western blot was used to detect the expression of mitogen activated protein kinase(MAPK)and nuclear factor-κB pathway related molecules.Results Compared with the hypoxia/reoxygenation group,the viability of H9C2 rat cardiomyocytes was increased in tea polyphenols group(P<0.05).Compared with the hypoxia/reoxygenation group,the content of malondialdehyde was decreased,while the content of superoxide dismutase,catalase and glutathione peroxidase were increased in the tea polyphenol group(P<0.05).Compared with the hypoxia/reoxygenation group,the contents of myeloperoxidase,IL-1β,TNF-αand IL-6 were decreased in the tea polyphenol group(P<0.05).Compared with the hypoxia/reoxygenation group,the expression of mitogen activated protein kinase and nuclear factor-κB pathway related molecules in tea polyphenol group was inhibited(P<0.05).Conclusion Tea polyphenols protect H9C2 rat cardiomyocytes from hypoxia/reoxygenation injury via mitogen activated protein kinase and nuclear factor-κB pathway.