PTEN编码蛋白对TGF-β_1刺激大鼠肾成纤维细胞增殖的抑制作用

Studies on the inhibitory effect of PTEN protein on the proliferation of rat renal fibroblasts stimulated by TGF-β_1

目的观察PTEN基因编码蛋白对TGF-β1刺激所致大鼠肾成纤维细胞增殖的影响。方法用重组PTEN腺病毒转染体外培养的大鼠肾成纤维细胞,倒置荧光显微镜检测绿色荧光蛋白表达,RT-PCR检测PTENmRNA表达。转染48h后TGF-β1体外刺激,TGF-β1刺激24h后MTT法检测细胞增殖活力,免疫细胞化学检测PCNA蛋白表达。结果PTEN腺病毒转染后绿色荧光蛋白和PTENmRNA表达明显增多。PTEN基因编码蛋白明显抑制TGF-β1刺激所致大鼠肾成纤维细胞增殖,并显著减少PCNA的表达。结论PTEN能抑制TGF-β1刺激所致大鼠肾成纤维细胞增殖,可能具有抑制残肾纤维化、延缓残肾毁损的作用。

Objective To investigate the effect of PTEN protein on the proliferation of rat renal fibroblasts resulted from the stimulation of TGF-β1. Methods In vitro, the cultured rat renal fibroblasts were transfected with the adenovirus containing PTEN. Invert fluorescent microscope was used to detect the GFP expression, meanwhile PTEN mRNA was determined by RT-PCR method, 48h after the transfection, TGF-β1 was added into the culture medium in a concentration of 10ng/ml. After another 24h, the proliferation rate of rat renal fibroblasts was determined by means of MTT method. Immunocytochemistry was also used to detect the expression of PCNA. Results As GFP positive fibroblasts were detected, the expression of PTEN mRNA was significantly increased in the rat renal fibroblasts which were transfected with the adenovirus containing PTEN. Accompanied with the expression of PTEN, the proliferation of cultured rat renal fibroblasts resulted from the stimulation of TGF-β1 was obviously inhibited, and the expression of PCNA was significantly decreased. Conclusion PTEN could suppress the proliferation of rat renal fibroblasts caused by stimulation of TGF-β1, and it might be useful to inhibit renal fibrosis and delay the rate of renal impairment.