缺氧缺糖诱导心肌细胞损伤中Notch信号对HIF-α及自噬相关基因Beclin1,LC3Ⅰ,LC3Ⅱ表达的影响  

Effects of Notch signal on the expressions of HIF-α and autophagyrelated genes Beclin1,LC3Ⅰ,LC3Ⅱ in oxygen-glucose deprivation induced myocardial cell injury

作　　者：孔令宇[1] 席錾[2] 马文婷 杨飞云[1] 牛丽丹[1] 石金河[1] KONG Ling-yu;XI Zan;MA Wen-ting;YANG Fei-yun;NIU Li-dan;SHI Jin-he(Department of Emergency,the First Affiliated Hospital of Xinxiang Medical University,Weihui 453100;Department of Pediatric Surgery,the First Affiliated Hospital of Xinxiang Medical University,Weihui 453100;Department of Diagnostic Laboratory of Sanquan College,Xinxiang Medical University,Xinxiang 453003,China)

机构地区：[1]新乡医学院第一附属医院急诊科,河南卫辉453100 [2]新乡医学院第一附属医院儿外科,河南卫辉453100 [3]新乡医学院三全学院诊断学实验室,河南新乡453003

出　　处：《中国应用生理学杂志》2019年第2期165-168,共4页Chinese Journal of Applied Physiology

基　　金：国家自然科学基金青年基金(81600677)

摘　　要：目的:探讨在缺氧缺糖诱导心肌细胞损伤的模型中Notch信号对低氧诱导因子(HIF-1α)及自噬相关的基因Beclin1,LC3Ⅰ,LC3Ⅱ的影响。方法:利用低氧培养箱与低糖DMEM培养基建立缺氧缺糖细胞(OGD)模型,细胞分为正常对照组,缺氧缺糖组(OGD group),缺氧缺糖+NC siRNA组(OGD+NC siRNA group),缺氧缺糖+Notch1 siRNA组(OGD+Notch1 siRNA group),缺氧缺糖+HIF-1αsiRNA组(OGD+HIF-1αsiRNA group),利用Western blot检测Notch1 siRNA与HIF-1αsiRNA的干预效果;利用Western blot检测Notch1 siRNA对模型细胞中HIF-1α表达的影响;利用CCK-8实验检测Notch1 siRNA与HIF-1αsiRNA对心肌细胞活性的影响;利用Western blot检测Notch1 siRNA与HIF-1αsiRNA对自噬相关的基因Beclin1,LC3Ⅰ,LC3Ⅱ的影响。结果:HIF-1αsiRNA可有效敲低模型心肌细胞HIF-1α的表达,而Notch1 siRNA可有效敲低模型心肌细胞中Notch1与HIF-1α的表达;Notch1 siRNA与HIF-1αsiRNA可降低缺氧缺糖细胞模型中心肌细胞的活性,且二者的作用之间没有统计学差异(P>0.05);Western blot结果显示Notch1 siRNA与HIF-1αsiRNA可降低模型细胞中自噬相关的基因Beclin1,LC3Ⅰ,LC3Ⅱ的表达,降低LC3Ⅱ/LC3Ⅰ的比率。结论:Notch1通过正向调节模型细胞HIF-1α的表达,进而提高缺氧缺糖诱导的自噬,发挥对心肌的保护作用。Objective: To investigate the effects of Notch signal on hypoxic induction factor( HIF-1α) and autophagy-associated genes Beclin1,LC3Ⅰ,LC3Ⅱ in oxygen-glucose deprivation( OGD) induced myocardial cell injury. Methods: The OGD model was established using hypoxic culture box and hypoglycemic DMEM medium. The cells were divided into normal control group,OGD group,OGD + NC siRNA group,OGD + Notch1 siRNA group and OGD + HIF-1α siRNA group. Western blot was used to detect the interference effects of HIF-1α siRNA and Notch1 siRNA. The effects of Notch1 siRNA and HIF-1α siRNA on the activity of myocardial cells in OGD model were detected by the CCK-8 assay. The effects of Notch1 siRNA and HIF-1α siRNA on autophage-associated genes Beclin1,LC3Ⅰ and LC3Ⅱ expression were detected by Western blot. Results: The results of Western blot showed that HIF-1α siRNA could effectively knock down the expression of HIF-1α in myocardial cells in OGD model,and Notch1 siRNA could effectively knock down the expression of Notch1 and HIF-1α in myocardial cells in OGD model. The result of CCK-8 assay showed that Notch1 siRNA and HIF-1α siRNA reduced the activity of myocardial cells in OGD model,and there was no statistical difference between the two groups.Western blot results showed that Notch1 siRNA and HIF-1α siRNA could reduce the expressions of the autophagy-associated genes Beclin1,LC3Ⅰ and LC3Ⅱ,and reduce the ratio of LC3Ⅱ to LC3Ⅰ at mRNA level. Conclusion: Notch1 plays a role in myocardial protection by regulating the expression of HIF-1α to regulate the autophagy in OGD model cells.

关 键 词：Notch信号通路 低氧诱导因子 缺血缺氧 自噬 

分 类 号：R459.9[医药卫生—治疗学]