SIRT1对TGF-β1活化肾脏成纤维细胞的抑制机制  

Inhibitory mechanism of SIRT1 on the activation of renal fibroblasts by transforming growth factorβ1

作　　者：盛梦丹 李金珂 周江桥[1] 陈玲[1] 邱涛[1] 唐艳[1] 张龙[1] SHENG Meng-dan;LI Jin-ke;ZHOU Jiang-qiao;CHEN Ling;QIU Tao;TANG Yan;ZHANG Long(Department of Organ Transplantation,People's Hospital of Wuhan University,Wuhan 430060,China)

机构地区：[1]武汉大学人民医院器官移植科,湖北武汉430060

出　　处：《生物技术》2022年第5期622-629,共8页Biotechnology

摘　　要：[目的]探讨沉默信息调节因子1(SIRT1)对转化生长因子β1(TGF-β1)活化肾脏成纤维细胞的抑制作用及机制。[方法]设立成纤维细胞NRK-49F组、TGF-β1刺激组(10 ngl/mL)、SIRT1蛋白低、中、高剂量组(10.0μg/mL、100.0μg/mL、1000.0μg/mL),以上各组每孔设6个平行样,培养72 h。采用MTT法测定各组细胞活力,流式细胞术测定细胞凋亡水平,ELISA法测定细胞纤维化程度α-SMA、FN、Vimentin蛋白水平,RT-PCR法及蛋白印迹法测定Smad3、SOSC1 mRNA和蛋白水平。[结果]TGF-β1刺激组OD值、存活率、α-SMA、FN、Vimentin蛋白、Smad3、SOSC1 mRNA和蛋白表达水平高于成纤维细胞NRK-49F组,凋亡率低于成纤维细胞NRK-49F组(P<0.05)。SIRT1蛋白低、中、高剂量组OD值、存活率、α-SMA、FN、Vimentin蛋白、Smad3、SOSC1 mRNA、Smad3、SOSC1蛋白表达水平低于TGF-β1刺激组,凋亡率高于TGF-β1刺激组,随着SIRT1蛋白剂量逐渐增加,SIRT1蛋白各剂量组OD值、存活率、α-SMA、FN、Vimentin蛋白、Smad3、SOSC1 mRNA和蛋白表达明显降低,凋亡率明显升高(P<0.05)。[结论]SIRT1蛋白通过抑制TGF-β1活化肾脏成纤维细胞中Smad3、SOSC1 mRNA与蛋白的表达,降低TGF-β1活化肾脏成纤维细胞的增殖及纤维化,促进细胞凋亡。[Objective]To investigate the inhibitory effect and mechanism of silence information regulator 1(SIRT1)on the activation of renal fibroblasts by transforming growth factorβ1.[Method]Fibroblast NRK-49F group,TGF-β1 stimulation group(10 ngl/mL),SIRT1 protein low,medium,and high dose groups(10.0μg/mL,100.0μg/mL,1000.0μg/mL)were established with 6 parallel samples in the above each groups and culture for 72 hours.MTT method was used to determine cell viability in each group,flow cytometry used to determine the level of apoptosis,ELISA method used to determine the degree of cell fibrosisα-SMA,FN,and Vimentin protein levels,and RT-PCR and Western Blot methods were used to determine Smad3,SOSC1 mRNA and protein level.[Result]The OD value,survival rate,α-SMA,FN,Vimentin protein,Smad3,SOSC1 mRNA and protein expression levels in TGF-β1 stimulation group were higher than those in fibroblast NRK-49F group,and the apoptosis rate was lower than that in fibroblasts cell NRK-49F group(P<0.05).The OD values of SIRT1 protein low,medium and high dose groups,survival rate,α-SMA,FN,Vimentin protein,Smad3,SOSC1 mRNA,Smad3,SOSC1 protein expression level was lower than those of TGF-β1 stimulation group,apoptosis rate higher than that in the TGF-β1 stimulation group(P<0.05).The OD value,survival rate,α-SMA,FN,Vimentin protein,Smad3,SOSC1 mRNA and protein expressions in each dose group were significantly decreased,and the apoptosis rate was significantly increased(P<0.05).[Conclusion]SIRT1 protein inhibits the expression of Smad3 and SOSC1 mRNA and protein in TGF-β1-activated renal fibroblasts,then reduces the proliferation and fibrosis of TGF-β1-activated renal fibroblasts,and promotes cell apoptosis.

关 键 词：沉默信息调节因子1 转化生长因子Β1 肾脏成纤维细胞 细胞增殖 

分 类 号：R692.9[医药卫生—泌尿科学]