miR-148b基因对高糖诱导的肾小球系膜细胞增殖、凋亡及免疫抑制因子的影响研究  被引量:3

Effect of miR-148b gene on proliferation,apoptosis and expression of immuno-suppressive factor TGF-β1 in glomerular mesangial cells induced by high glucose

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作  者:汤夏莲[1] 陈平[1] 唐义平[1] 张敏[1] 曹晓红[1] TANG Xia-Lian;CHEN Ping;TANG Yi-Ping;ZHANG Min;CAO Xiao-Hong(Department of Geriatric Endocrinology,Sichuan Provincial People忆s Hospital,Sichuan Academy of Medical Sciences,Chengdu 610072,China)

机构地区:[1]四川省医学科学院.四川省人民医院老年内分泌科,成都610072

出  处:《中国免疫学杂志》2018年第12期1794-1797,1803,共5页Chinese Journal of Immunology

摘  要:目的:探讨miR-148b基因对高糖诱导的肾小球系膜细胞增殖、凋亡及免疫抑制因子TGF-β1表达的影响。方法:小鼠肾小球系膜细胞(MMCs)分三组处理,即低糖+miR-148b scramble组(LG组)、高糖+miR-148b scramble组(HG组)和高糖+miR-148b inhibitor组(HG+miR-148b inhibitor组),其中miR-148b scramble和miR-148b inhibitor的转染参照脂质体LipofectamineTM2000转染说明进行瞬时转染,收集转染48 h的细胞,通过qRT-PCR检测各组细胞中miR-148b的mRNA表达;CCK8及流式细胞仪分别检测细胞活力及凋亡率;Western blot检测免疫抑制因子TGF-β1及NF-κB信号通路p-IκBα及下游相关基因cyclin D1和Bax的蛋白表达。结果:高糖可上调miR-148b的表达,转染miR-148b inhibitor后miR-148b的表达明显降低;与LG组比较,HG组细胞活力显著升高,细胞凋亡率显著升高,TGF-β1、p-IκBα、cyclin D1和Bax的蛋白表达均明显升高,与HG组比较,HG+miR-148b inhibitor组细胞活力显著降低,细胞凋亡率显著降低,TGF-β1、p-IκBα、cyclin D1和Bax的蛋白表达均明显降低(P<0. 05)。结论:抑制肾小球系膜细胞miR-148b基因表达可抑制高糖诱导的细胞活力、凋亡率及免疫抑制因子TGF-β1表达的增加,机制可能与下调NF-κB信号通路有关。Objective:To investigate the effect of miR-148b gene on the proliferation,apoptosis and the expression of immuno-suppressive factor TGF-β1in glomerular mesangial cells induced by high glucose.Methods:Mouse glomerular mesangial cells were divided into three groups,low sugar+miR-148b scramble group(LG group),high glucose+miR-148b scramble group(HG group)and high glucose+miR-148b inhibitor group(HG+miR-148b inhibitor group),and miR-148b scramble and miR-148b inhibitor was transient transfected into MMCs according to the transfection of liposome LipofectamineTM2000.Cells were transfected for48h,miR-148b mRNA expression was detected by qRT-PCR;CCK8and flow cytometry were used to detect cell viability and apoptosis rate;the expression of TGF-β1,p-IκBα,cyclinD1and Bax protein were detected by Western blot.Results:High glucose could increase the expression of miR-148b,and the expression of miR-148b decreased significantly after transfection of miR-148b inhibitor.Compared with LG group,cell viability increased significantly in HG group,apoptosis rate was significantly increased,the expression of TGF-β1,p-IκBα,cyclinD1and Bax protein were significantly increased;compared with HG group,cell viability decreased significantly in HG+miR-148b group,the apoptosis rate was significantly decreased,the expression of TGF-β1,p-IκBα,cyclinD1and Bax protein were decreased significantly(P<0.05).Conclusion:Inhibiting of miR-148b gene expression in glomerular mesangial cells induced by high glucose can inhibit the increase of cell viability,apoptosis rate and TGF-β1expression,and the mechanism may be related to the down regulation of NF-κB signaling pathway.

关 键 词:miR-148b基因 肾小球系膜细胞 高糖 凋亡 免疫 

分 类 号:R587.2[医药卫生—内分泌]

 

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