Mn_(3)O_(4)纳米酶处理对缺氧/复氧心肌细胞HIF-1α/BNIP3信号通路的影响

Mn_(3)O_(4)nanoenzyme treatment alleviates hypoxia/reoxygenation-induced cardiomyocyte injury via HIF-1α/BNIP3 signaling pathway

目的:探讨四氧化三锰(Mn_(3)O_(4))纳米酶处理对缺氧/复氧(H/R)心肌细胞低氧诱导因子-1α(HIF-1α)/B淋巴细胞瘤-2/腺病毒E1B 19kDa相互作用蛋白3(BNIP3)信号通路的影响。方法:建立H9C2大鼠心肌细胞H/R损伤模型,将H9C2大鼠心肌细胞随机分为对照组、模型组、四氧化三锰纳米颗粒(Mn_(3)O_(4)NP)组、二甲氧基雌二醇(2-ME2)组和Mn_(3)O_(4)NP+2-ME2组。CCK-8法检测细胞活力。生化法检测丙二醛(MDA)、乳酸脱氢酶(LDH)水平。Western blot和实时荧光定量PCR分别检测HIF-1α、BNIP3、苄氯素1(Beclin-1)、家蚕隔离体蛋白1(P62)的蛋白质和mRNA表达水平。结果:与对照组相比,模型组心肌细胞活力下降,LDH及MDA表达水平升高,HIF-1α、BNIP3、Beclin-1蛋白及mRNA表达水平升高,P62表达水平降低(P均<0.05)。与模型组相比,Mn_(3)O_(4)NP组心肌细胞活力上升,LDH及MDA表达水平降低,HIF-1α、BNIP3、Beclin-1蛋白及mRNA表达水平升高,P62表达水平降低(P均<0.05),2-ME2组变化相反(P均<0.05)。与Mn_(3)O_(4)NP+2-ME2组相比,Mn_(3)O_(4)NP组心肌细胞活力上升,LDH及MDA表达水平降低,HIF-1α、BNIP3、Beclin-1表达水平升高,P62表达水平降低(P均<0.05),2-ME2组变化相反(P均<0.05)。结论:Mn_(3)O_(4)NP可能通过激活HIF-1α/BNIP3信号通路促进H9C2心肌细胞自噬,从而减轻H/R诱导的心肌损伤。

Objective:To explore the effect of Mn_(3)O_(4)nanoenzyme treatment on HIF-1α/BNIP3 signaling pathway in hypoxia/reoxygenation(H/R)cardiomyocytes.Methods:After establishing a H/R injury model using H9C2 mice,cardiomyocytes were randomly divided into five groups:control group,model group,Mn_(3)O_(4)NP group,2-ME2 group and Mn_(3)O_(4)NP+2-ME2 group.In each group,CCK-8 technique was used to determine myocardial cell viability,and Western blot and qRT-PCR were used to detect the protein and mRNA expression levels of HIF-1α,BNIP3,Beclin-1,P62,respectively.MDA and LDH levels were assessed by biochemical methods.Results:Compared with the control group,myocardial cell viability was decreased,LDH and MDA levels were increased,HIF-1α,BNIP3,Beclin-1 protein and mRNA expression levels were increased,and P62 expression was reduced in the model group(P<0.05).Compared with the model group,myocardial cell viability was increased,LDH and MDA levels were decreased,HIF-1α,BNIP3 and Beclin-1 expression levels were increased,and P62 expression was decreased in the Mn_(3)O_(4)NP group(P<0.05).Opposite changes occurred in the 2-ME2 group(P<0.05).Compared with the Mn_(3)O_(4)NP+2-ME2 group,myocardial cell viability was increased,LDH and MDA levels were decreased,HIF-1α,BNIP3 and Beclin-1 expression levels were increased,and P62 expression was reduced in the Mn_(3)O_(4)NP group(P<0.05).Opposite changes were detected in the 2-ME2 group(P<0.05).Conclusion:Mn_(3)O_(4)NP may promote autophagy of H9C2 cardiomyocytes by activating HIF-1α/BNIP3 signaling pathway,thereby alleviating H/R-induced myocardial injury.