尿酸对人尿道成纤维细胞TGF-β1,Smad3的影响.doc

PAGE PAGE 1 尿酸对人尿道成纤维细胞TGF-β1、Smad3的影响 郭 旭，卢根生 (第三军医大学西南医院全军泌尿外科研究所，重庆 400038) 摘　要：目的　观察人尿道成纤维细胞在尿液、尿酸刺激下TGF-β1、Smad3的表达变化，探讨尿道瘢痕的形成机制。方法 体外培养人尿道成纤维细胞，以尿液、350 μmol/L尿酸浓度的尿酸刺激溶液刺激人尿道成纤维细胞，通过免疫荧光染色和逆转录-聚合酶链反应（RT-PCR）分别检测各组细胞TGF-β1、Smad3因子表达变化, ELISA方法比较各组细胞上清液Ⅰ型胶原蛋白差异。结果 免疫荧光检测提示TGF-β1与Smad3蛋白在人尿道成纤维细胞中均有表达，尿液及生理浓度的尿酸刺激后，TGF-β1与Smad3的mRNA表达上调、蛋白表达明显升高（P﹤0.05）；ELISA法检测显示，尿液和尿酸刺激人尿道成纤维细胞24、48h后，细胞上清液中Ⅰ型胶原蛋白含量显著升高( P﹤0.05)。结论 尿酸刺激引起人尿道成纤维细胞TGF-β1、Smad3的表达升高；尿酸可能对尿道瘢痕的形成起促进作用。 关键词:尿道瘢痕；成纤维细胞；转化生长因子-β1；尿液；尿酸 中图法分类号： 文献标识码：A Effects of uric acid on TGF-β1 and SMAD3 expression in human urethral fibroblasts GUO Xu,Lu GEN-sheng (Institute of Urinary Surgery Research, Southwest Hospital, Third Military Medical University, Chongqing 400038, China) Abstract: Objective To observe the effects of urine on TGF-β1 and Smad3 expression in cultured human urethral fibroblasts and explore the mechanism of urethral scar fibrosis. Methods The human urethral fibroblasts were cultured and stimulated with urine or 350μmol/L uric acid. The mRNA and protein expression of TGF-β1 and SMAD3 were measured by PT-PCR and immuno-fluorescent staining, respectively. Collagen Ⅰ in the supernatant of different groups was detected and compared by ELISA. Results TGF-β1 and SMAD3 could be detected in human urethral fibroblasts by immuno-fluorescent staining. After stimulated by urine or physiological concentration of uric acid,both the protein and mRNA expression of TGF-β1 and SMAD3 in human urethral fibroblasts were increased significantly（P﹤0.05）. By ELISA,the upregulation of Collagen I in the supernatant was also discovered 24 or 48 hours later after the stimulation of urine or uric acid. Conclusion The stimulation of uric acid could increase the expression of TGF-β1 and SMAD3. Uric acid probably take part in promotion of urethral scar fibrosis. Key words: urethral scar; fibroblast; TGF-β1；urine; uric acid 作者简介:郭 旭,男,河北省黄骅人,硕士研究生,医师,主要从事尿道瘢痕方面的研究。电话: (023) 687