球形脂联素对间歇低氧所致H9C2细胞损伤的保护作用及机制
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E?mail:dybbin@163.com

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R563.9

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国家自然科学基金项目(81700090,81770086)


Effects and mechanisms of globular adiponectin on intermittent hypoxia induced H9C2 cell injury
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    摘要:

    目的:探讨Parkin介导的线粒体自噬在球形脂联素(globular adiponectin,gAPN)减轻间歇低氧(intermittent hypoxia, IH)所致H9C2心肌细胞损伤中的作用及相关机制。方法:建立H9C2细胞IH模型来模拟阻塞性睡眠呼吸暂停低通气综合征。 采用流式细胞术检测细胞凋亡。采用JC-1试剂盒检测线粒体膜电位(mitochondrial membrane potential,MMP)。采用免疫印迹法检测Parkin、Bax、Bcl-2蛋白表达。采用Parkin siRNA抑制Parkin基因表达。采用LC3与线粒体红色荧光探针进行荧光共定位检测线粒体自噬。结果:IH暴露后,H9C2细胞凋亡率、Bax及Parkin蛋白表达水平、线粒体自噬明显升高,MMP下降,Bcl-2 蛋白表达水平下降;给予gAPN保护后,IH+gAPN组细胞凋亡率、Bax蛋白表达水平较IH组下降,MMP较IH组升高,Bcl-2及 Parkin蛋白表达水平、线粒体自噬也较IH组升高。抑制Parkin基因表达后,线粒体自噬明显下降,线粒体损伤、心肌细胞凋亡率明显增加。结论:gAPN通过上调Parkin介导的线粒体自噬减轻IH所致H9C2心肌细胞损伤。

    Abstract:

    Objective:This study aims to evaluate the effect of Parkin mediated mitophagy on globular adiponectin(gAPN) ameliorated H9C2 cardiomyocytes injury induced by intermittent hypoxia(IH)and the potential mechanisms. Methods:The cell IH model was established. H9C2 cell apoptosis was detected by flow cytometry. The mitophagy was detected by fluorescence colocalization of LC3 and mitochondrial red marker. The protein expression levels of Bax,Bcl-2 and Parkin were detected by Western blot. The mitochondrial membrane potential(MMP) was detected by JC-1 kit. Results:After IH exposure,the apoptosis of H9C2 cardiomyocytes,protein expression levels of Bax and Parkin,mitophagy were increased,MMP and Bcl-2 protein expression were decreased. After gAPN treatment,the apoptosis of H9C2 cardiomyocytes,Bax protein expression level were decreased,the MMP, protein expression levels of Bcl-2 and Parkin,and mitophagy were increased. Inhibiting Parkin gene expression decreased the mitophagy,and increased the mitochondrial injury and H9C2 cell apoptosis. Conclusion:gAPN ameliorated the IH induced H9C2 cardiomyocytes injury through upregulating Parkin mediated mitophagy.

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丁文筱,张蔷,丁宁,张希龙,董艳彬.球形脂联素对间歇低氧所致H9C2细胞损伤的保护作用及机制[J].南京医科大学学报(自然科学版),2022,42(11):1523-1529,1538

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  • 在线发布日期: 2022-11-24
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