胺碘酮促进小鼠HL1心房肌细胞热处理诱导的凋亡、炎症及氧化应激

Amiodarone promotes heat-induced apoptosis,inflammation and oxidative stress in mouse HL1 atrial myocytes

目的探讨热处理对心房肌细胞的损伤作用及胺碘酮对心房肌细胞热处理损伤的影响。方法先测定不同温度和不同胺碘酮浓度下的细胞活力,确定最适合的加热温度和胺碘酮浓度,将细胞分为空白对照组、热处理组、胺碘酮组,空白对照组将体外培养小鼠HL1心房肌细胞置于37℃恒温水浴箱,热处理组将心房肌细胞置于高温水浴箱,胺碘酮组将加入胺碘酮溶液的心房肌细胞培养皿置于高温水浴箱。通过MTS法检测细胞活力,通过流式细胞术检测细胞凋亡,利用酶联免疫法检测白介素-6(IL-6)、白介素-1β(IL-1β)和肿瘤坏死因子-α(TNF-α)水平,通过分光光度计检测丙二醛(MDA)、超氧化物歧化酶(SOD)水平,观察热处理后心房肌细胞损伤类型及胺碘酮对热处理诱导的心房肌细胞凋亡、炎症、氧化应激水平的影响。结果与空白组比较,热处理后心房肌细胞存活率与SOD活性减少(P<0.001),热处理组的IL-1β及MDA水平增加(P<0.01)、凋亡率及IL-6增加极其显著(P<0.001);与热处理组相比,胺碘酮组心房肌细胞存活率降低(P<0.01)、SOD活性下降(P<0.05)、凋亡率增加(P<0.05),而IL-1β、IL-6及MDA水平增加(P<0.01),TNF-α水平增加显著(P<0.001)。结论热处理可导致小鼠HL1心房肌细胞发生凋亡、炎症、氧化应激导致细胞损伤,胺碘酮溶液可促进热处理诱导后的细胞凋亡、炎症、氧化应激反应从而进一步加重小鼠HL1心房肌细胞损伤。

Objective To investigate the injury types of atrial myocytes induced by heat exposure and the effect of amiodarone on heat-induced injuries in atrial myocytes.Methods The optimal temperature for heat exposure and optimal concentration of amiodarone were determined by measuring the cell viability exposed to different temperatures and different concentrations of amiodarone.Heat exposure of HL1 atrial myocytes was conducted using a water bath,and the effect of amiodarone on cell viability was assessed with MTS method;cell apoptosis was detected using flow cytometry,and the levels of IL-1β,IL-6,TNF-α,SOD and MDA were detected with enzyme-linked immunosorbent assay(ELISA).Results Compared with the blank control cells,the cells exposed to a temperature of 52℃showed a significantly decreased survival rate and a lowered SOD activity(P<0.001)with increased IL-1βand MDA levels(P<0.01)and markedly increased apoptosis rate and IL-6 level(P<0.001).Compared with the heat exposure group,amiodarone resulted in significantly decreased survival rate of the atrial myocytes(P<0.01),obviously decreased SOD activity(P<0.05),and increased cell apoptosis rate(P<0.05)and IL-1β,IL-6,MDA and TNF-αlevels(P<0.01 or 0.001).Conclusion Heat exposure induces apoptosis,inflammation and oxidative stress in mouse HL1 atrial myocytes,and amiodarone can enhance the effects of heat exposure to aggravate the cell injuries.