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[Notch信号对氧糖剥夺诱导的心肌细胞损伤中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 autophagy- related genes Beclin1, LC3I, LC3II in oxygen-glucose deprivation induced myocardial cell injury].

作者信息

Kong Ling Yu, Xi Zan, Ma Wen Ting, Yang Fei Yun, Niu Li Dan, Shi Jin He

机构信息

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.

出版信息

Zhongguo Ying Yong Sheng Li Xue Za Zhi. 2019 Feb;35(2):165-168. doi: 10.12047/j.cjap.5729.2019.036.

Abstract

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.

摘要

目的

探讨Notch信号对氧糖剥夺(OGD)诱导的心肌细胞损伤中缺氧诱导因子(HIF-1α)及自噬相关基因Beclin1、LC3I、LC3II的影响。

方法

采用缺氧培养箱和低糖DMEM培养基建立OGD模型。将细胞分为正常对照组、OGD组、OGD + NC siRNA组、OGD + Notch1 siRNA组和OGD + HIF-1α siRNA组。采用蛋白质免疫印迹法检测HIF-1α siRNA和Notch1 siRNA的干扰效果。采用CCK-8法检测Notch1 siRNA和HIF-1α siRNA对OGD模型中心肌细胞活性的影响。采用蛋白质免疫印迹法检测Notch1 siRNA和HIF-1α siRNA对自噬相关基因Beclin1、LC3I和LC3II表达的影响。

结果

蛋白质免疫印迹结果显示,HIF-1α siRNA可有效敲低OGD模型中心肌细胞HIF-1α的表达,Notch1 siRNA可有效敲低OGD模型中心肌细胞Notch1和HIF-1α的表达。CCK-8法检测结果显示,Notch1 siRNA和HIF-1α siRNA降低了OGD模型中心肌细胞的活性,两组间差异无统计学意义。蛋白质免疫印迹结果显示,Notch1 siRNA和HIF-1α siRNA可降低自噬相关基因Beclin1、LC3I和LC3II的表达,并在mRNA水平降低LC3II与LC3I的比值。

结论

在OGD模型细胞中,Notch1通过调节HIF-1α的表达来调控自噬,从而发挥心肌保护作用。

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