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中国应用生理学杂志 ›› 2019, Vol. 35 ›› Issue (2): 165-168.doi: 10.12047/j.cjap.5729.2019.036

• 研究论文 • 上一篇    下一篇

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*

孔令宇1,席錾2,马文婷3,杨飞云1,牛丽丹1,石金河1△   

  1. 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
  • 出版日期:2019-03-28 发布日期:2019-06-27
  • 通讯作者: Tel: 13525074567; E-mail: sjh4402901@163.com
  • 基金资助:
    国家自然科学基金青年基金(81600677)

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

KONG Ling-yu 1, XI Zan 2, MA Wen-ting 3, YANG Fei-yun 1, NIU Li-dan 1, SHI Jin-he1△   

  1. 1. Department of Emergency, the First Affiliated Hospital of Xinxiang Medical University, Weihui 453100;
    2. Department of Pediatric Surgery, the First Affiliated Hospital of Xinxiang Medical University, Weihui 453100;
    3. Department of Diagnostic Laboratory of Sanquan College, Xinxiang Medical University, Xinxiang 453003, China
  • Online:2019-03-28 Published:2019-06-27

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-1α siRNA组(OGD + HIF-1α siRNA group),利用Western blot检测Notch1 siRNA与HIF-1α siRNA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-1α siRNA对心肌细胞活性的影响;利用Western blot检测Notch1 siRNA与HIF-1α siRNA对自噬相关的基因Beclin1,LC3I,LC3II的影响。结果: HIF-1α siRNA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-1α siRNA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-1α siRNA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。

关键词: Notch信号通路, 低氧诱导因子, 缺血缺氧, 自噬

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.

Key words: Notch signaling pathway, hypoxic induction factor, ischemia and hypoxia, autophagy

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