PTEN编码蛋白对TGF-β_1刺激大鼠肾成纤维细胞分泌ColⅣ、FN的抑制作用  

The inhibitive effect of PTEN protein on secretion of collagen Ⅳ and fibronectin by stimulation of by TGF-β_1 in rat

作　　者：程悦[1] 王代红[1] 张耀全[1] 李芙蓉[1] 袁发焕[1] 

机构地区：[1]第三军医大学新桥医院肾内科全军肾脏病中心重庆市肾脏病研究所,重庆400037

出　　处：《解放军医学杂志》2007年第7期719-721,共3页Medical Journal of Chinese People's Liberation Army

摘　　要：目的观察PTEN基因编码蛋白对转化生长因子β1(TGF-β1)刺激大鼠肾成纤维细胞分泌Ⅳ型胶原(ColⅣ)、纤维连接蛋白(FN)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,实验分为4组,对照组,TGF-β1组(给予TGF-β1刺激),PTEN+TGF-β1组(Ad-PTEN转染后给予TGF-β1刺激)和GFP+TGF-β1组(Ad-GFP转染后给予TGF-β1刺激)。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ELISA法检测细胞上清中ColⅣ、FN水平。结果PTEN腺病毒转染后可见明显绿色荧光蛋白表达,PTEN mRNA表达增多。TGF-β1组及GFP+TGF-β1组较对照组ColⅣ、FN分泌量明显增多,PTEN+TGF-β1组较TGF-β1组及GFP+TGF-β1组ColⅣ、FN分泌量明显降低(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分泌ColⅣ、FN,故可能具有抑制残肾纤维化、延缓残肾毁损速度的作用。Objective To investigate the role of PTEN protein in secretion of collagen Ⅳ and fibronectin after stimulation of transforming growth factor beta 1 （TGF-β1） from renal fibroblasts of rat in vitro. Methods The cultured rat renal fibroblasts were transfected with the reconstructed adenovirus containing PTEN or adenovirus only containing green fluorescence protein （GFP）. The fibroblsts were treated in four manners： control group with no added treatment, TGF-β1 group with TGF-β1, stimulation, PTEN＋ TGF -β1group with TGF-β1 stimulation after Ad-PTEN transfection, and GFP＋ TGF-β1 group with TGF-β1 stimulation after Ad-GFP transfection. Invert fluorescent microscope was used to detect the GFP expression, meanwhile the PTEN mRNA was determined by RT-PCR method. 36h after the transfection, TGF-β1 was added into the culture medium in a concentration of 10ng/ml. After another 24h, ELISA method was used to evaluate the level of collagen Ⅳ and fibronectin. Results The expressions of both GFP and PTEN mRNA increased obviously after the rats＇ renal fibroblasts were transfected with adenovirus. The secretion of collagen Ⅳ and fibronectin increased significantly in both TGF-β1 group and GFP＋ TGF-β1 group compared with that in control group, and decreased markedly in PTEN＋ TGF-β1 group compared with that in both TGF-β1 group and GFP＋ TGF-β1 group （P〈0. 05）. When TGF-β1 was applied to the cultured rats＇ renal fibroblasts, the secretion of collagen Ⅳ and fibronectin was increased obviously, and it could be inhibited by the overexpression of PTEN protein （P〈0. 05）. Conclusion PTEN protein could inhibit the secretion of collagen Ⅳ and fibronectin in rats＇ renal fibroblasts stimulated by TGF-β1, implying that PTEN protein might be useful in restraining fibrosis in the remaining kidney and delaying impairment of renal function.

关 键 词：基因 PTEN 转化生长因子β1 胶原Ⅳ型 成纤维细胞 纤连蛋白类 肾 

分 类 号：R692.39[医药卫生—泌尿科学]