lncRNA GAS5对人心脏微血管内皮细胞缺血再灌注损伤的影响及机制实验研究OA

Objective:To investigate the effect of long non-coding RNA(lncRNA)growth arrest-specific tran-script 5(GAS5)on ischemia-reperfusion(IR)injury in human cardiac microvascular endothelial cells(HCMEC)and its possible mechanism.Methods:HCMECs were divided into the following groups:normal control group(routine culture without intervention),model group(IR injury model established),negative control group(IR+transfection with negative control lentivirus),GAS5 overexpression group(IR+transfection with lncRNA GAS5 overexpression lentivirus),and GAS5 knockdown group(IR+transfection with lncRNA GAS5 interference lentivirus).Morphologi-cal characteristics of cells in each group were observed.RT-qPCR was performed to detect GAS5 mRNA levels.ELISA was used to measure inflammatory cytokine levels.MTT assay was conducted to evaluate cell proliferation.TUNEL assay was employed to assess apoptosis.Western blot was used to detect the protein expression of LC3 and Beclin1.Results:HCMECs in the normal control group exhibited tight adherence,regular morphology,uniform cyto-plasm,and no cell detachment.In contrast,cells in the model group demonstrated significantly reduced adherence,shrinkage and rounding,increased cytoplasmic granularity,and partial detachment from the culture dish.No obvious morphological differences were observed between the negative control group and the model group.These IR-related morphological alterations were markedly ameliorated in the GAS5 overexpression group,whereas the GAS5 knock-down group exhibited further aggravated cellular damage compared with the negative control group.Compared with the normal control group,the model group showed increased apoptosis rate and elevated levels of interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α),along with decreased cell proliferation rate,GAS5 mRNA expression,and pro-tein expression of LC3 and Beclin1(all P＜0.05).No statistically significant differences were observed between the model group and the negative control group in these parameters(all P＞0.05).The GAS5 overexpression group demonstrated improved changes in the above indicators compared with the negative control group,while the GAS5 knockdown group showed aggravated changes(all P＜0.05).Conclusion:Upregulation of lncRNA GAS5 expression can attenuate IR-induced HCMEC injury,and the mechanism may be associated with the activation of autophagy.