The effect of LncRNA-NEAT1-mediated miR-182-5p expression regulation on lupus nephritis mesangial cell injury
ZHANG Lu-lu
XIE Rui
LIAO Zhi-min
WU Gang
WAN Bo
SUN Wei
ZHOU Lian-hong
Abstract：Objective To investigate the effect of long non-coding RNA(LncRNA)nuclear paraspeckle assem-bly transcript 1(NEAT1)on the damage of renal mesangial cells in lupus nephritis(LN)through micro RNA(miR)-182-5p/forkhead box protein O1(FoxO1)/β-catenin axis.Methods The peripheral blood of LN patients(LN group)and physical examination of healthy persons(healthy control group)who were admitted to the Second People's Hospital of Liangshan Prefecture from March 2019 to July 2021 were collected.Real time fluorescence quantitative poly-merase chain reaction(qRT-PCR)method was used to assess the expression levels of miR-182-5p,FoxO1,β-cate-nin mRNA in peripheral blood mononuclear cells.Enzyme linked immunosorbent assay(ELISA)method was used to measured serum inflammatory factors tumor necrosis factor-α(TNF-α),interleukin(IL)-6 and IL-1β contents.The LN renal mesangial cell model was constructed by treating 20％serum of LN patients with kidney mesangial cells.Af-ter modeling,the cells were divided into control group(normal culture,no transfection),model group(cultured in 5μg/mL lipopolysaccharide),si-NC group(transfected with si-NC,and cultured in 5 μg/mL lipopolysaccharide),si-NEAT1 group(transfected with si-NEAT1,and cultured in 5 μg/mL lipopolysaccharide),si-NEAT1+anti-miR-NC group(cotransfected with si-NEAT1 and anti-miR-NC,and cultured in 5 μg/mL lipopolysaccharide)and si-NEAT1+anti-miR-182-5p group(cotransfected with si-NEAT1 and anti-miR-182-5p,and cultured in 5 μg/mL lipopolysaccharide).The qRT-PCR method was used to detect the expression levels of NEAT1 and miR-182-5p of cells in each group.ELISA method was used to determine the levels of cellinflammatory factors in each group.Lactate de-hydrogenase(LDH)kit was used to detect the LDH content of cells in each group.Cell counting kit 8(CCK-8)experi-ment was used to detect cell proliferation activity in each group.The flow cytometry was used to analyze cell apoptosis in each group.Western blotting was used to assess the expression levels of NEAT1,FoxO1 and β-catenin proteins in each group.Results Compared with the healthy control group,the expression levels of NEAT1,FoxO1,β-catenin mRNA in peripheral blood mononuclear cells and serum inflammatory factors TNF-α,IL-6,IL-1β levels in the LN group were significantly increased,while the expression level of miR-182-5p was significantly reduced(P＜0.05).Compared with the control group,the renal mesangial cell proliferation activity,Fox01 and β-catenin protein levels,and LDH,TNF-α,IL-6 and IL-1β levels in the model group were significantly increased,and the expression level of miR-182-5p was significantly decreased(P＜0.05).After knocking down NEAT1,the cell proliferation activity and inflam-matory response were significantly weakened,and FoxO1 and β-catenin protein expression levels were significantly down-regulated(P＜0.05).Inhibition of miR-182-5p expression could reduce the effects of NEAT1 knockdown on cell pro-liferation,inflammatory factors and FoxO1 and β-catenin protein expression in LN renal mesangial cell model(P＜0.05).There was no significant change in the apoptotic rate among cells in each group(P＞0.05).Conclusion Knockdown of NEAT1 may inhibit excessive proliferation of LN mesangial cells and reduce inflammation by negatively reg-ulating miR-182-5p expression.This effect may be related to the inhibition of the FoxO1/β-catenin pathway.
Keywords：nuclear paraspeckle assembly transcript 1lupus nephritismiR-182-5p/forkhead box protein O1/β-catenin axisrenal mesangial cells
Publication Date：2024-01-28
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：7( 99-105 )