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转化生长因子-β1诱导小鼠足细胞凋亡实验研究
引用本文:王宇晖,童孟立,杨汝春.转化生长因子-β1诱导小鼠足细胞凋亡实验研究[J].浙江医学,2013,35(19):1718-1721.
作者姓名:王宇晖  童孟立  杨汝春
作者单位:杭州市中医院肾内科,310007
基金项目:浙江省中医药管理局立项资助课题(2006C108)
摘    要:目的探讨转化生长因子-β1(TGF-β1)是否能诱导小鼠足细胞株凋亡以及诱导凋亡的途径。方法(1)体外培养小鼠足细胞株;(2)应用不同浓度(10^-1~104ng/L)TGF-β1诱导小鼠足细胞凋亡;(3)10^3ng/LTGF-β1诱导不同时间(12、24、48h)后观察足细胞凋亡情况;(4)采用流式细胞仪检测AnnexinV、Caspase3;(5)实时定量PCR及Western印迹法检测p38丝裂原活化蛋白激酶(p38MAPK)、Smad2、Smad3、Smad7mRNA及蛋白质的表达情况。结果(1)小鼠足细胞在不同浓度(10^-1~10^4ng/L)TGF-β1刺激下,细胞凋亡率明显高于正常对照组(均P〈0.05);(2)随着TGF-β1浓度增加、凋亡时间增加,足细胞凋亡率增高(P〈0.05);p38MAPK、Smad2、Smad3、Smad7mRNA表达增加;TGF-β1 103ng/L为最佳诱导小鼠足细胞凋亡浓度,24h为最佳诱导凋亡时间;(3)与正常对照组比较,10^3ng/LTGF-β1诱导后Caspase3阳性细胞比率显著增加(P〈001).p38MAPK、Smad2、Smad3、Smad7蛋白质表达亦显著增加(P〈0.01)。结论TGF-β1能诱导小鼠足细胞凋亡,呈浓度及时间依赖性。TGF-β1通过Smad通路、p38MAPK通路以及线粒体通路诱导小鼠足细胞凋亡。

关 键 词:TGF-β1,足细胞凋亡

TGF-β1 induces apoptosis of mouse podocytes in vitro
WANG Yuhui,TONG Mengli,YANG Ruchun.TGF-β1 induces apoptosis of mouse podocytes in vitro[J].Zhejiang Medical Journal,2013,35(19):1718-1721.
Authors:WANG Yuhui  TONG Mengli  YANG Ruchun
Institution:. Department of Nephrology, Hangzhou Hospital of Traditional Chinese Medicine, Hangzhou 310007, China
Abstract:Objective To investigate the effect of TGF-β1 on apoptosis of mouse podocytes and its signal pathway. Methods Cultured mouse podocytes were treated with different concentrations of TGF-β1(0,1x10-1N104 ng/L) for 24 h, or treated with 103 ng/L TGF-β1 for different time duration (12, 24, 48h). Annexin V and Caspase 3 were measured by flow cytometry. The mRNAs and proteins of P38MAPK, Smad2, Smad3 and Smad7 were detected by real time PCR and Western blotting. Results Compared to controls(0ng/L TGF- 15 1) the ratio of apoptotic podocytes and mRNA expressions of p38MAPK, Smad2, Smad3 and Smad7 were increased in TGF-β1 (10-1-104ng/L) treatment groups in a concentration and time-dependent manner (P〈0.05). Treatment with TGF--β1 (103rig/L) for 24 h induced maximal apoptosis in podocytes. The ratio of Caspase3-positive podocytes was increased after treated with TGF--β1(103ng/L, 24h) compared to controls(P〈0.01). Conclusion TGF-β1induces the apop- tosis of mouse podocytes time-dependently and dose-dependently, in which Smad, p38MAPK signal pathway and mitochondri- on might be involved.
Keywords:TGF-β1  Podocyte Apoptosis
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