PEA3在白蛋白诱导肾小管上皮细胞损伤中的作用

Role of PEA3 in the injury of renal proximal tubular cells induced by albumin

目的:探讨PEA3在白蛋白诱导的肾近端小管上皮细胞损伤中的作用及机制。方法:体外培养小鼠近端肾小管上皮细胞,实验组分为对照组、空载对照组、PEA3过表达组、白蛋白造模组和PEA3处理组。实时荧光定量PCR检测PEA3转染效率。应用Annexin V-FITC/PI双染和Hoechst染色检测细胞凋亡。Western blot法和RT-PCR检测肾小管上皮细胞标志物E-cadherin、α-SMA及Vimentin的蛋白和m RNA变化水平。结果:1PEA3质粒转染后,过表达组的PEA3表达水平增高了2.5倍;2加入白蛋白刺激后,与对照组相比小管细胞凋亡增加,过表达PEA3显著抑制白蛋白诱导的小管细胞凋亡;3白蛋白刺激降低E-cadherin的表达,而增加了α-SMA和Vimentin的表达,PEA3过表达阻断白蛋白对E-cadherin的抑制作用,并降低α-SMA和Vimentin的表达。结论:PEA3可抑制白蛋白诱导的小管上皮细胞凋亡和上皮-间充质转分化。

Objective：To investigate the role of PEA3 in albumin induced mouse proximal tubular cells（m PTCs）apoptosis and its underlying mechanism. Methods：The cells were cultured with albumin in vitro,and divided into the control group,the vehicle control group,the PEA3 overexpression group,the albumin model group and the PEA3 overexpression in albumin model group. Real-time fluorescence quantitative polymerase chain reaction was used to investigate the level of PEA3 gene transfection efficiency. Flow cytometry and Hoechst staining technique were performed to analyze the apoptosis of m PTCs. The m RNA and protein levels of Ecadherin,α-SMA and Vimentin were detected by RT-PCR and Western bolt. Results：Transfection of PEA3 over-expresed the m RNA level of PEA3 for 2.5 fold. The apoptosis of m PTCs treated with albumin was increased compared with the control group,and overexpression of PEA3 ameliorated albumin-induced apoptosis. Albumin treatment reduced the expression of E-cadherin and increased the expression of Vimentin and α-SMA. Overexpression of PEA3 reversed the reduction of E-cadherin and ameliorated the increase of Vimentin and α-SMA. Conclusion：Overexpression of PEA3 can protect m PTCs from apoptosis and epithelialmesenchymal transition induced by albumin.