MiR-145-5p基因对LPS诱导的肾小球系膜细胞凋亡及细胞外基质分泌的研究  

Effects of miR-145-5p gene on apoptosis and extracellular matrix secretion of glomerular mesangial cells induced by LPS

作　　者：周瑛[1] 沈伟兴 夏宇苗 王丽[1] 蒋妙华[1] 郑永健[1] 傅鹏[1] ZHOU Ying;SHEN Wei-xing;XIA Yu-miao;WANG Li;JIANG Miao-hua;ZHENG Yong-jian;FU Peng(Department of Nephrology,East City Hospital of Shanghai Yangpu District,Shanghai 200438,China)

机构地区：[1]上海市杨浦区市东医院肾内科,上海200438

出　　处：《武警后勤学院学报（医学版）》2018年第5期374-378,共5页Journal of Logistics University of PAP（Medical Sciences）

基　　金：上海市杨浦区医学重点学科项目(YP16ZB03)

摘　　要：【目的】探讨上调microRNA-145-5p(miR-145-5p)基因表达对脂多糖(lipopolysaccharide,LPS)诱导的肾小球系膜细胞凋亡及细胞外基质分泌的影响。【方法】大鼠肾小球系膜细胞株HBZY-1随机分为空白组、LPS组和LPS+miR-145-5p mimics组(miR-145-5p组),miR-145-5p mimics转染参照脂质体LipofectamineTM2000说明,刺激24 h后,MTT及流式细胞术分别检测细胞活力及凋亡率;Western blotting检测纤维连结蛋白(fibronectin,FN)、转化生长因子-β1(transforming growth factor-β1,TGF-β1)、B细胞淋巴瘤/白血病-2(B cell lymphoma/lewkmia-2,Bcl-2)、Bcl-2相关X蛋白(Bcl-2 associated X protein,Bax)的蛋白表达。【结果】转染miR-145-5p mimics后的HBZY-1细胞miR-145-5p表达明显高于空白组(P<0.05)。与空白组比较,LPS组细胞活力及FN、TGF-β1和Bcl-2的蛋白表达显著升高,细胞凋亡率及Bax的蛋白表达显著降低(P<0.05)。与LPS组比较,miR-145-5p组细胞活力及FN、TGF-β1和Bcl-2的蛋白表达显著降低,细胞凋亡率及Bax的蛋白表达显著升高(P<0.05)。【结论】上调miR-145-5p基因表达可抑制LPS诱导的肾小球系膜细胞活力,通过下调FN和TGF-β1表达抑制细胞外基质分泌,通过下调Bcl-2和上调Bax表达诱导细胞凋亡。[Objective] To explore the effects of of miR-145-5p gene up-regulation on apoptosis and extracellular matrix secretion of glomerular mesangial cells induced by lipopolysaccharide （LPS）. [ Methods ] Rat glomerular mesangial cell line HBZY-1 was selected and the cells were randomly divided into blank group, LPS group and LPS ＋ miR-145-5p mimics group （miR-145-5p group, ）. Cells are transfected referring to LipofeetamineTM2000, and cells were stimulated for 24 h. The cell viability and apoptosis rate were detected by MTT and flow eytometry, and Western blotting was used to detect the protein expressions of fibronectin （FN）, transforming growth factor- β1 （TGF- β1）, B cell lymphoma/lewkmia-2 （Bcl-2） and Bcl-2 associated X protein （Bax）. [Results] The expression of miR-145-5p in HBZY-1 cells transfected with miR- 145-5p mimics was significantly higher than that in the blank group （P〈0.05）. Compared with the blank group, the cell vitality and expressions of FN, TGF- β1 and Bcl- 2 significantly increased in the LPS group, and the apoptosis rate and protein expression of Bax decreased significantly （P〈0.05）. Compared with the LPS group, the cell vitality and expressions of FN, TGF-β1 and Bcl-2 significantly decreased in the miR-145-Sp group, and the apoptosis rate and expression of Bax protein were significantly increased （P〈0.05）. [Conclusion ] Up-regulation of miR-145-5p gene expression can inhibit the activity of glomerular mesangial cells induced by LPS. Thesecretion of extracellular matrix can be inhibited by the down- regulation of FN and TGF- 131 expression, and apoptosis can be induced by the down-regulation of Bcl-2 and up-regulation of Bax.

关 键 词：肾小球系膜细胞 miR-145-5p基因 脂多糖诱导 凋亡 细胞外基质分泌 

分 类 号：R692[医药卫生—泌尿科学]