霉酚酸酯对狼疮性肾炎小鼠肾小球系膜细胞增殖、炎性反应的影响及分子机制研究  

Effect of mycophenolate mofetil on proliferation and inflammatory response of glomerular mesangial cells in lupus nephritis mice and its molecular mechanism

作　　者：李均[1] 杨晓娟[1] 冉阳 肖骏[1] 郝培 LI Jun;YANG Xiaojuan;RAN Yang;XIAO Jun;HAO Pei(Chongqing Emergency Medical Center,Chongqing 400014,China)

机构地区：[1]重庆市急救医疗中心重庆市第四人民医院,重庆400014 [2]青岛市城阳区人民医院,山东青岛266109

出　　处：《陕西医学杂志》2023年第8期973-976,991,共5页Shaanxi Medical Journal

基　　金：重庆市自然科学基金资助面上项目(CSTC2021JCYJ-MSXMX1061)。

摘　　要：目的:探究霉酚酸酯(MMF)抑制狼疮性肾炎(LN)肾小球系膜细胞(GMC)增殖及炎性反应的分子机制。方法:分离、培养LN细胞,将细胞分为:对照组,LN组,0.01μmol/L MMF组,0.1μmol/L MMF组,1.0μmol/L MMF组,转染miR-NC+LN组,转染miR-455-3p+LN组,转染anti-miR-NC+0.1μmol/L MMF组,转染anti-miR-455-3p+0.1μmol/L MMF组。用实时荧光定量PCR(RT-qPCR)法检测各组细胞中miR-455-3p的相对表达水平;用CCK-8法、流式细胞术、酶联免疫法(ELISA)、蛋白免疫印迹法(Western blot)分别检测各组细胞的增殖、凋亡率及肿瘤坏死因子α(TNF-α)、白介素1β(IL-1β)水平,活化型半胱氨酸天冬氨酸蛋白酶-3(Cleaved caspase-3)蛋白表达水平。结果:与对照组比较,LN组miR-455-3p相对表达水平、细胞凋亡率以及Cleaved caspase-3蛋白表达水平显著降低(均P<0.05),细胞增殖,TNF-α、IL-1β表达水平显著升高(均P<0.05);与LN组比较,0.01μmol/L MMF组、0.1μmol/L MMF组以及1.0μmol/L MMF组细胞增殖和炎症因子表达水平降低(均P<0.05),miR-455-3p、细胞凋亡率以及Cleaved caspase-3蛋白表达水平升高(均P<0.05),miR-455-3p+LN组与miR-NC+LN比较,细胞增殖和炎症因子表达水平降低(均P<0.05),miR-455-3p表达,凋亡率以及Cleaved caspase-3蛋白水平显著升高(均P<0.05);与anti-miR-NC+0.1μmol/L MMF组比较,anti-miR-455-3p+0.1μmol/L MMF组miR-455-3p表达,凋亡率及Cleaved caspase-3蛋白表达显著降低,细胞增殖和炎症因子表达水平显著升高(均P<0.05)。结论:MMF能够抑制LN肾小球系膜细胞的增殖和炎性反应、促进凋亡、改善LN,这一作用与miR-455-3p的表达调控有关。Objective:To explore the molecular mechanism of mycophenolate mofetil(MMF)inhibiting the proliferation and inflammatory response of glomerular mesangial cells(GMC)in lupus nephritis(LC).Methods:The isolated and cultured MCs of LN mice were divided into control group,LN model group,0.01μmol/L MMF group,0.1μmol/L MMF group,1.0μmol/L MMF group,miR-NC+LN group,miR-455-3p+LN group,anti-miR-NC+0.1μmol/L MMF group,anti-miR-455-3p+0.1μmol/L MMF group.The relative expression levels of miR-455-3p were detected by RT-qPCR.CCK-8 method,flow cytometry,ELISA,Western blot were used to detect the proliferation,apoptosis rate,the expression levels of TNF-α,IL-1βand Cleaved caspase-3 protein were analyzed.Results:Compared with the control group,the relative expression level of miR-455-3p,apoptosis rate and the expression level of Cleaved caspase-3 protein in the LN model group were significantly decreased,the cell proliferation and the levels of TNF-αand IL-1βwere significantly increased(all P<0.05).Compared with the LN model group,the 0.01μmol/L MMF group,0.1μmol/L MMF group and 1.0μmol/L MMF group had decreased cell proliferation and inflammation level,and the expression level of miR-455-3p,apoptosis rate and the expression level of Cleaved caspase-3 protein increased(all P<0.05).Compared with miR-NC+LN group,the cell proliferation and the expression levels of inflammatory factors in miR-455-3p+LN group were decreased,while the expression of miR-455-3p,apoptosis rate and Cleaved caspase-3 protein level were significantly increased(all P<0.05).Compared with the anti-miR-NC+0.1μmol/L MMF group,the anti-miR-455-3p+0.1μmol/L MMF group showed significantly lower miR-455-3p expression,apoptosis rate and Cleaved caspase-3 protein expression,while the cell proliferation and inflammation levels were significantly increased(all P<0.05).Conclusion:Mycophenolate mofetil can inhibit the proliferation and inflammatory reaction of LN glomerular mesangial cells,promote apoptosis,and improve LN,which is related to the regulation of

关 键 词：狼疮性肾炎 肾小球系膜细胞 霉酚酸酯 miR-455-3p 细胞增殖 细胞凋亡 炎性反应 

分 类 号：R593.24[医药卫生—内科学]