Effect of LncRNA KCNQ1OT1 on high glucose-induced human retinal microvascular endothelial cell injury by regulating the miR-7-5p/YAP1 axis

WANG Weijing; WANG Kai

doi:10.13431/j.cnki.immunol.j.20250121

Abstract：Objective To discuss whether LncRNA KCNQ1OT1 can regulate the miR-7-5p/Yes-associated protein 1(YAP1)axis and affect the high glucose-induced human retinal microvascular endothelial cells(HRMECs)injury.Methods HRMECs were divided into the control group,the high glucose group,the KCNQ1OT1 negative control group,the KCNQ1OT1 interference group,the KCNQ1OT1 interference+miRNA inhibition negative control group,the KCNQ1OT1 interference+miR-7-5p inhibition group,and the KCNQ1OT1 interference+YAP1 empty vector group,and KCNQ1OT1 interference+YAP1 overexpression group.The control group was treated with 5.5 mmol/L glucose for 48 h,while the high glucose group was treated with 25.0 mmol/L glucose for 48 h.For the other cells that needed transfection,the corresponding plasmids were first transfected into each group of cells using Lipofectamine 2000 transfection reagent,and then the cells were treated with 25.0 mmol/L glucose for 48 h.The expression levels of KCNQ1OT1,miR-7-5p,and YAP1 mRNA were detected in each group,as well as the levels of lactate dehydrogenase(LDH),tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),interleukin-1β(IL-1β),reactive oxygen species(ROS),and malondialdehyde(MDA),the activities of superoxide dismutase(SOD)and catalase(CAT),the status of apoptosis,and the protein expression levels of Bcl-2-associated X protein(Bax),B-cell lymphoma-2(Bcl-2),Cleaved caspase-3,and YAP1 in each group.Additionally,to validate the targeting relationship between miR-7-5p and KCNQ1OT1/YAP1 was verified.Results Compared with the control group,the high-glucose group showed increased mRNA expression levels of KCNQ1OT1 and YAP1;elevated levels of TNF-α,IL-6,IL-1β,LDH,MDA,and ROS,as well as a higher apoptosis rate;increased protein expression levels of Bax,Cleaved caspase-3,and YAP1;decreased expression of miR-7-5p and reduced Bcl-2 protein expression;and decreased activities of CAT and SOD(P<0.05).Interfering with the expression of KCNQ1OT1 could improve the above index levels in HRMECs(P<0.05).Inhibition of miR-7-5p expression or overexpression of YAP1 could both weaken the improvement effect of silencing KCNQ1OT1 expression on HRMECs injury induced by high glucose(P<0.05).Conclusion Silencing the expression of KCNQ1OT1 may alleviate high glucose-induced HRMECs injury by regulating the miR-7-5p/YAP1 axis.