MiR-181a-5p regulates the proliferation and apoptosis of glomerular mesangial cells in lupus nephritis mice through HMGB1/NF-κB signaling pathway

YANG Xiaofang; JIA Xinyan; FENG Wenjun

doi:10.11958/20253169

Abstract：Objective To explore the regulatory mechanism of microRNA(miR)-181a-5p on the proliferation and apoptosis of glomerular mesangial cells in mice with lupus nephritis(LN)through the high mobility group protein box 1(HMGB1)/nuclear factor kappa B(NF-κB)signaling pathway.Methods LN mouse mesangial cells were isolated and cultured in vitro,and divided into the blank group,the mimics-NC group,the miR-181a-5p mimics group,the Si-NC group,the Si-HMGB1 group(transfected with Si-HMGB1),the mimics-NC+pcDNA-NC group,the miR-181a-5p mimics+pcDNA-NC group,the mimics-NC+pcDNA-HMGB1 group and the miR-181a-5p mimics+pcDNA-HMGB1 group.MRL/MPJ mouse mesangial cells were isolated and cultured in vitro as the control group.qRT-PCR method was implemented to measure the mRNA expression levels of miR-181a-5p and HMGB1 in cells.Dual-luciferase experiments were performed to verify the targeting relationship of miR-181a-5p to HMGB1.CCK-8 method and flow cytometry were implemented to measure cell proliferation and apoptosis,respectively.Western blot assay was performed to measure the protein expression levels of HMGB1,human inhibitor of nuclear factor κB-α(IκB-α)and NF-κB p65 in cells.Results There was a targeting relationship between miR-181a-5p and HMGB1.Compared with the control group,the cell proliferation rate,HMGB1 and NF-κB p65 protein expression were obviously increased in the blank group,and the miR-181a-5p and IκB-α expression were obviously decreased(P＜0.05).Compared with the mimics-NC group,the cell proliferation rate,HMGB1 and NF-κB p65 protein expression were obviously decreased in the miR-181a-5p mimics group,and the miR-181a-5p and IκB-α expression were obviously increased(P＜0.05).Compared with the Si-NC group,the cell proliferation rate,HMGB1 and NF-κB p65 protein expression were obviously decreased in the Si-HMGB1 group,and the IκB-α expression was obviously increased(P＜0.05).The overexpression of HMGB1 was able to reverse the inhibitory effect of up-regulated miR-181a-5p on the excessive proliferation of glomerular mesangial cells in LN mice(P＜0.05).Conclusion Overexpression of miR-181a-5p can inhibit the excessive proliferation of glomerular mesangial cells in LN mice,which may be related to the inhibition of HMGB1/NF-κB signaling pathway.