中国临床药理学与治疗学2023，Vol.28Issue(12)：1331-1338,8.DOI:10.12092/j.issn.1009-2501.2023.12.002
大黄素通过miR-96-5p靶向叉头蛋白K2减轻狼疮性肾炎肾小球系膜细胞的损伤
Emodin reduces the injury of glomerular mesangial cells in lupus ne-phritis by targeting forkhead protein K2 through miR-96-5p
摘要
Abstract
AIM:To investigate the injury of emo-din(EMO)in reduce of glomerular mesangial cells(MCs)in lupus nephritis by targeting forkhead pro-tein K2(FOXK2)through miR-96-5p.METHODS:The contents of 24 h urine protein,serum urea ni-trogen(BUN)and serum creatinine(Scr)in MRL/faslpr mice(lupus nephritis group)and MRL/MPJ mice(control group)were detected.MCs were sep-arated,purified and divided into:MCs group(MCs without any treatment),L-EMO group(MCs treated with 10 μmol/L Emodin),M-EMO group(MCs treat-ed with 25 μmol/L Emodin),H-EMO group(MCs treated with 50 μmol/L Emodin),H-EMO+miR-96-5p-NC group(MCs treated with 50 μmol/L Emodin and transfected with miR-96-5p-NC),and H-EMO+miR-96-5p-minic group(MCs treated with 50 μmol/L Emodin and transfected with miR-96-5p-minic).Double luciferase report experiment was used to verify the targeting relationship between miR-96-5p and FOXK2.The real-time quantitative fluores-cent polymerase chain reaction(qRT-PCR)was used to detect the expression of miR-96-5p.West-ern blot was used to detect the expression of FOXK2 and apoptosis related proteins.The enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of inflammatory factors in MCs.cell count kit 8(CCK-8)was used to determine the activity of MCs.Annexin-V FITC/PI double staining was used to detect apoptosis of MCs.RESULTS:Compared with the control group,24 h urinary pro-tein content,serum BUN and Scr levels in the lupus nephritis group were significantly increased(P＜0.05).Compared with the MCs group,the miR-96-5p expression,interleukin1β(IL-1β),interleukin6(IL-6),tumor necrosis factor-α(TNF-α),A450 value and B-lymphoblastoma-2(Bcl-2)protein in the L-EMO group,M-EMO group and H-EMO group were significantly decreased(P＜0.05),the FOXK2 level,cell apoptosis rate,Bcl-2 related X gene(Bax),as-partate specific cysteine proteinase-3(cleaved Cas-pase-3)protein levels were significantly increased,respectively(P＜0.05),the effect of Emodin was dose-dependent.Compared with the H-EMO group and H-EMO+miR-96-5p-NC group,H-EMO+miR-96-5p-minic group obviously increased the miR-96-5p expression,inflammatory factor levels,A450 value and Bcl-2 protein level(P＜0.05),and obviously de-creased FOXK2 level and cell apoptosis rate(P＜0.05).CONCLUSION:EMO can reduce the injury of lupus nephritis MCs by down-regulating miR-96-5p and then up-regulating FOXK2.关键词
大黄素/miR-96-5p/叉头蛋白K2/狼疮性肾炎/肾小球系膜细胞Key words
emodin/miR-96-5p/forkhead box protein K2/lupus nephritis/glomerular mesangial cells分类
医药卫生引用本文复制引用
施珊红,林威远,张杰群,郑燕玲..大黄素通过miR-96-5p靶向叉头蛋白K2减轻狼疮性肾炎肾小球系膜细胞的损伤[J].中国临床药理学与治疗学,2023,28(12):1331-1338,8.基金项目
福建省卫生健康科技计划项目(2022G0184) （2022G0184）
