| 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 |
| DOI： |
| 分类号:R575.2 |
| 基金项目:浙江省中医药管理局立项资助课题（2006C108） |
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| TGF-β1 induces apoptosis of mouse podocytes in vitro |
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WANG Yuhui,TONG Mengli,YANG Ruchun
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. Department of Nephrology, Hangzhou Hospital of Traditional Chinese Medicine, Hangzhou 310007, China
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| 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. |
| Key words: TGF-β1, Podocyte Apoptosis |
