低氧诱导心肌细胞损伤中，低氧诱导因子-1α（HIF-1α）与长链非编码GAS5之间的相互作用调控JAK1/STAT3信号通路。
The interplay between HIF-1α and long noncoding GAS5 regulates the JAK1/STAT3 signalling pathway in hypoxia-induced injury in myocardial cells.
作者信息
Li Yanwei, Song Bing, Liu Jinlei, Li Yuqiang, Wang Jiebing, Liu Na, Cui Wei
机构信息
Management Center of Chronic Diseases, The First Affiliated Hospital of Jinzhou Medical University, Jinzhou, China.
Department of Endocrine and Metabolic Diseases, The First Affiliated Hospital of Jinzhou Medical University, Jinzhou, China.
出版信息
Cardiovasc Diagn Ther. 2021 Apr;11(2):422-434. doi: 10.21037/cdt-20-773.
BACKGROUND
Long non-coding RNA (lncRNA) GAS5 is associated with hypoxia-induced diseases whereas hypoxia-inducible factor-1α (HIF-1α) plays an important role in hypoxic injury of cells. The current study explores the regulatory functions of GAS5/HIF-1α which co-play in anoxic injury among rat cardiomyocytes H9C2 cells.
METHODS
Hypoxia model was established through anoxic incubation while normal culture of H9C2 cells was considered as control. The expression levels of GAS5 and HIF-1α were quantified through RT-qPCR. CCK-8 was applied to determine cell viability. Cell apoptosis rate was calculated using flow cytometry whereas inflammatory cytokines were detected using ELISA method. The impact of downregulating GAS5 or HIF-1α or both upon hypoxic cells was assessed on the basis of changes in cell viability, apoptosis, and inflammatory response. The activity of JAK1/STAT3 signaling was evaluated through RT-qPCR for mRNA expression. AG490 was introduced to inactivate JAK1/STAT3 pathway and to unveil the impact of JAK1/STAT3 signaling on GAS5/HIF-1α and cell viability, apoptosis and inflammation in hypoxic cells.
RESULTS
The results infer that hypoxia suppressed cell viability, promoted inflammation and apoptosis among H9C2 cells. GAS5 or HIF-1α recorded higher expression in hypoxia-induced cells whereas the cell viability got restored with reduction in inflammation and apoptosis. The downregulation of HIF-1α enhanced the protective effect of knocking down GAS5 in hypoxia H9C2 cells. JAK1/STAT3 signaling pathway got activated in hypoxic cells and was regulated by GAS5 and HIF-1α. The inhibition of signaling pathway increased the cell viability but it decreased both inflammation and apoptosis.
CONCLUSIONS
GAS5 and HIF-1α could regulate hypoxic injury in H9C2 cells through JAK1/STAT3 signaling pathway. This scenario suggests that the inhibitors of GAS5 and HIF-1α may synergize with AG-490 to protect myocardial cells from hypoxic injury.
背景
长链非编码RNA（lncRNA）GAS5与缺氧诱导的疾病相关，而缺氧诱导因子-1α（HIF-1α）在细胞缺氧损伤中起重要作用。本研究探讨GAS5/HIF-1α在大鼠心肌细胞H9C2细胞缺氧损伤中共同发挥的调节功能。
方法
通过缺氧孵育建立缺氧模型，将H9C2细胞的正常培养作为对照。通过RT-qPCR定量GAS5和HIF-1α的表达水平。应用CCK-8测定细胞活力。使用流式细胞术计算细胞凋亡率，而使用ELISA方法检测炎性细胞因子。基于细胞活力、凋亡和炎症反应的变化，评估下调GAS5或HIF-1α或两者对缺氧细胞的影响。通过RT-qPCR检测mRNA表达来评估JAK1/STAT3信号的活性。引入AG490以失活JAK1/STAT3途径，并揭示JAK1/STAT3信号对缺氧细胞中GAS5/HIF-1α以及细胞活力、凋亡和炎症的影响。
结果
结果表明，缺氧抑制了H9C2细胞的活力，促进了炎症和凋亡。GAS5或HIF-1α在缺氧诱导的细胞中表达较高，而细胞活力随着炎症和凋亡的减少而恢复。HIF-1α的下调增强了在缺氧H9C2细胞中敲低GAS5的保护作用。JAK1/STAT3信号通路在缺氧细胞中被激活，并受GAS5和HIF-1α调节。信号通路的抑制增加了细胞活力，但降低了炎症和凋亡。
结论
GAS5和HIF-1α可通过JAK1/STAT3信号通路调节H9C2细胞的缺氧损伤。这表明GAS5和HIF-1α的抑制剂可能与AG-490协同作用，保护心肌细胞免受缺氧损伤。
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