PTEN编码蛋白对TGF-β1刺激大鼠肾成纤维细胞增殖和ColⅣ与FN分泌的抑制作用

目的：观察张力蛋白同源物基因（PTEN）编码蛋白对转化生长因子-β1（TGF-β1）刺激所致大鼠肾成纤维细胞增殖和Ⅳ型胶原（ColⅣ）、纤连蛋白（FN）分泌的影响。方法：用重组PTEN腺病毒转染体外培养大鼠肾成纤维细胞,倒置荧光显微镜检测绿色荧光蛋白表达,RT-PCR检测PTEN mRNA表达。转染36h后TGF-β1体外刺激,TGF-β1刺激24h后MTT法检测细胞增殖活力,免疫细胞化学法检测PCNA表达,ELISA法检测细胞上清中ColⅣ、FN水平。结果：PTEN腺病毒转染36h后可见明显绿色荧光蛋白表达,PTEN mRNA表达明显增多（P〈0.01）。PTEN腺病毒转染后给予TGF-β1刺激组较单纯TGF-β1刺激组平均光密度值（OD）显著降低（P〈0.01）,PCNA表达显著减少（P〈0.01）,且ColⅣ、FN的分泌水平明显下降（P〈0.05）。结论：PTEN基因编码蛋白能抑制TGF-β1刺激所致大鼠肾成纤维细胞增殖、PCNA表达及ColⅣ、FN分泌,可能具有抑制残肾纤维化、延缓残肾毁损速度的作用。

The Inhibitive Effect of PTEN Protein on Rat Renal Fibroblasts Proliferation and Collagen Ⅳ and Fibronectin Expression Induced by TGF-β1

Objective：To investigate the inhibitive effect of PTEN protein on rat renal fibroblasts proliferation and Collagen Ⅳ and Fibronectin expression induced by TGF-β1.Methods：The cultured rat renal fibroblasts were transfected with adenovirus containing PTEN,invert fluorescent microscope was used to detect the green fluorescent protein（GFP） expression,PTEN mRNA was measured by RT-PCR method. 36 h after transfection,TGF-β1 was added into the culture medium with a concentration of 10 ng/ml.The proliferation of rat renal fibroblasts was measured by means of methylthio tetrazole（MTT） method 24 h hours after transfection. Immunocytochemistry was used to detect the expression of PCNA. ELISA method was used to evaluate the level of collagen Ⅳ and fibronectin.Results：As GFP positive fibroblasts were detected,the expression of PTEN mRNA was significantly increased in transfected rat renal fibroblasts. Accompanied with the expression of PTEN, the proliferation of TGF-β1 stimulated rat renal fibroblasts was obviously inhibited,the expression of PCNA was significantly decreased and the level of collagen Ⅳ and fibronectin was significantly decreased.Conclusion：PTEN can suppress the proliferation and the expression of collagen Ⅳ and fibronectin in rat renal fibroblasts stimulated by TGF-β1, which might be useful to inhibit renal fibrosis and retard the progression of renal disease.