氧化白藜芦醇通过核因子E2相关因子2信号通路缓解心肌缺血再灌注损伤

Oxyresveratrol alleviates myocardial ischemia reperfusion injury through nuclear factor erythroid-2-related factor 2 signaling pathway

目的探讨氧化白藜芦醇(Oxy)在缺氧/复氧(H/R)诱导的心肌损伤中的保护作用。方法小鼠心肌HL-1细胞培养于DMEM高糖完全培养基作为Ctrl组;通过缺氧6 h/复氧2 h刺激HL-1细胞,建立心肌细胞H/R损伤模型,作为H/R组;Ctrl组细胞在20μM Oxy的DMEM高糖完全培养基预孵育2 h,后进行H/R刺激,作为H/R+Oxy组。通过细胞计数试剂盒-8检测细胞活力;采用蛋白免疫印迹技术检测B淋巴细胞瘤2(Bcl-2)、Bcl-2相关X蛋白(Bax)、核因子E2相关因子2(Nrf-2)蛋白表达水平;使用终端尿苷核苷酸末端标记法和烟酸己可碱染色法评估各组细胞凋亡情况;使用实时荧光定量PCR技术分析Nrf-2、血红素加氧酶1(Ho-1)、醌氧化还原酶1(Nqo-1)mRNA表达水平;采用超氧化物歧化酶(SOD)和丙二醛(MDA)试剂盒,利用酶标仪检测各组细胞内SOD和MDA水平。结果与Ctrl组比较,H/R组细胞活力降低、Bax蛋白升高、Bcl-2蛋白降低、Bax/Bcl-2升高、细胞凋亡数量升高、细胞内SOD水平降低、MDA水平升高,差异有统计学意义(P<0.05)。与H/R组比较,H/R+Oxy组细胞活力升高,Bax蛋白降低、Bcl-2蛋白升高、Bax/Bcl-2降低、细胞凋亡数量降低、细胞内SOD水平升高、MDA水平降低、Nrf-2蛋白升高、Nrf-2、Ho-1、Nqo-1 mRNA表达水平升高,差异有统计学意义(P<0.05)。结论Oxy可抑制H/R诱导的心肌细胞损伤,可能通过激活Nrf-2信号通路发挥抗氧化作用。

Objective To investigate the protective effects of oxyresveratrol(Oxy)on hypoxia/reoxygenation(H/R)-induced cardiac oxidative myocardial injury.Methods Mouse myocardial HL-1 cells were cultured in DMEM high-sugar complete medium as the control group(Ctrl group).HL-1 cells were subjected to 6 hours of hypoxia and 2 hours of reoxygenation to establish a myocardial H/R injury model(H/R group).Ctrl group cells were pre-incubated with 20μM Oxy in DMEM complete medium for 2 hours before H/R stimulation(H/R+Oxy group).Cell viability was detected by cell counting kit-8 assay.Protein expression levels of B-cell lymphoma-2(Bcl-2),Bcl-2-associated X(Bax),and nuclear factor erythroid-2-related factor 2(Nrf-2)were detected by Western blot.Apoptosis in each group was evaluated using terminal uridine nucleotide end labeling(TUNEL)and Hoechst assay.The mRNA expression levels of Nrf-2,heme oxygenase-1(Ho-1),and quinone oxidoreductase 1(Nqo-1)were analyzed using real-time quantitative PCR.Superoxide dismutase(SOD)and malondialdehyde(MDA)levels in cells from each group were measured using corresponding assay kits and a microplate reader.Results Compared to the Ctrl group,the H/R group showed decreased cell viability,increased Bax protein level,decreased Bcl-2 protein level,elevated apoptosis,reduced SOD level,and increased MDA level,with statistically significant differences(P<0.05).Compared to the H/R group,the H/R+Oxy group exhibited increased cell viability,decreased Bax protein level,increased Bcl-2 protein level,reduced apoptosis,increased SOD level,decreased MDA level,and elevated mRNA expression of Nrf-2,Ho-1,and Nqo-1,with statistically significant differences(P<0.05).Conclusion Oxy can inhibit H/R-induced myocardial cell injury,and may play an antioxidant role by activating the Nrf-2 signaling pathway.