The role of long non-coding RNA-MEG3 in ischemia-reperfusion injury of human cardiac microvascular endothelial cells
Objective To study the role of long non-coding RNA maternally expressed gene 3(lncRNA MEG3)in ischemia-reperfusion injury(IRI)of human cardiac microvascular endothelial cells(HCMECs).Methods HCMECs were treated with blank control,induced with hypoxia and reoxygenation(H/R),or induced with H/R followed by transfection of si-MEG3.Relative levels of lncRNA MEG3 and inflammatory factors,including the interleukin beta(IL-β),IL-6,IL-8,IL-10 and tumor necrosis factor alpha(TNF-α)were detected by quantitative real-time polymerase chain reaction(qRT-PCR).Cell proliferation and apoptosis were detected by CCK-8 assay and flow cytometry,respectively.Protein expressions of key proteins in the phosphatidylinositol 3-kinase/protein kinase B/endothelial nitric oxide synthase(PI3K/Akt/eNOS)signaling pathway were detected by Western blot.The enzyme-linked immunosorbent assay(ELISA)was performed to measure relative levels of nitric oxide(NO),superoxide dissolving enzyme(SOD),vascular endothelial growth factor(VEGF)and reactive oxygen species(ROS).Results qRT-PCR data showed that the relative level of lncRNA MEG3 in H/R-induced HCMECs was significantly higher than that of controls[6.87±0.239]times vs[1.00±0.026]times,n=3,t=42.32,P<0.0001),suggesting the role of lncRNA MEG3 in IRI of HCMECs.Cell proliferation was significantly inhibited in H/R-induced HCMECs than that of controls,which was further inhibited in H/R-induced HCMECs with knockdown of lncRNA MEG3(P<0.01).Protein expressions of p-PI3K,p-AKT and p-eNOS were significantly lower in H/R-induced HCMECs than those of controls,which were further downregulated in H/R-induced HCMECs with knockdown of lncRNA MEG3(P<0.05).Significantly decreased NO,SOD and VEGF levels and increased ROS level were detected in H/R-induced HCMECs than those of controls,which were more pronounced in H/R-induced HCMECs with knockdown of lncRNA MEG3(P<0.05).Significantly increased mRNA levels of inflammatory factors were detected in H/R-induced HCMECs than those of controls,which were significantly higher in H/R-induced HCMECs with knockdown of lncRNA MEG3(P<0.01).Conclusion LncRNA MEG3 exerts the protective effect on H/R of HCMECs.Overexpressing lncRNA MEG3 is a potential therapeutic target for alleviating IRI in cardiac microvascular vessels.
万方数据
维普
