收稿日期: 2017-12-26
网络出版日期: 2018-08-21
基金资助
新乡医学院科研培育基金（2014QN119）
The regulation of hypoxic induced factor on autophagy of myocardial cells in oxygen-glucose deprivation model
Received date: 2017-12-26
Online published: 2018-08-21
目的 探讨在缺血缺氧模型中低氧诱导因子（HIF-1α）对自噬相关基因Beclin1，LC3I，LC3II的影响及其对自噬的调节作用。 方法 建立缺血缺氧细胞模型，利用western blot 检测HIF-1α siRNA的干预效果；利用CCK-8实验检测HIF-1α siRNA对心肌细胞活性的影响；利用RT-qPCR与western blot检测HIF-1α siRNA对自噬相关基因Beclin1，LC3I，LC3II的影响。 结果 Western blot结果显示HIF-1α siRNA可有效敲低模型心肌细胞中HIF-1α的表达；CCK-8实验结果显示，HIF-1α siRNA可降低缺血缺氧细胞模型中心肌细胞的活性；RT-qPCR与western blot结果均提示HIF-1α siRNA可降低模型细胞中自噬相关基因Beclin1，LC3I，LC3II的表达，降低LC3II/LC3I的比率。 结论 HIF-1α siRNA敲低缺血缺氧细胞模型中的HIF-1α表达后，导致心肌细胞的活性进一步降低，细胞中Beclin1表达降低，LC3II/LC3I的比率降低，HIF-1α正调控缺血缺氧诱导的自噬而发挥心肌保护作用。
宋海岩 , 连辉 , 张毅敏 , 付升旗 . 低氧诱导因子对缺氧缺糖诱导的心肌细胞损伤模型中自噬的调控作用[J]. 中国临床解剖学杂志, 2018 , 36(4) : 419 -422 . DOI: 10.13418/j.issn.1001-165x.2018.04.013
Objective To investigate the effect of HIF-1α on autophage-associated genes Beclin1, LC3I and LC3II, and the regulation on autophagy. Methods The model of oxygen-glucose deprivation (OGD) was established, and western blot was used to detect interference effect of HIF-1α siRNA. The CCK-8 assay was used to detect the effect of HIF-1α siRNA on the activity of myocardial cells in OGD model. The effect of HIF-1α siRNA on autophagy-associated genes Beclin1, LC3I and LC3II expression was detected by RT-qPCR and 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. The result of CCK-8 assay showed that HIF-1α siRNA could reduce the activity of myocardial cells in OGD model. RT-qPCR and western blot results showed that HIF-1α siRNA could reduce the expression of the autophagy-associated genes Beclin1, LC3I and LC3II, and reduce the ratio of LC3II to LC3I at mRNA and protein level. Conclusion HIF-1α siRNA could knock down the expression of HIF-1α in the OGD model, resulting in further decrease of the activity of myocardial cells，with the Beclin1 expression and ratio of LC3II/LC3I reduced. HIF-1α plays a role of myocardial protection in the OGD model by regulation of autophagy.
Key words： Ischemia; Hypoxia; Autophagy; Hypoxic induction factor; Myocardial injury
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