雷帕霉素对大鼠尿道成纤维细胞增殖和胶原形成的影响  

Effects of rapamycin on the proliferation and collagen production of urethral fibroblasts in male rats

作　　者：李建平[1] 郑舜生[1] 刘莹[1] 李和程[1] 付德来[1] 翟晓强[1] 种铁[1] LI Jian-ping ZHENG Shun-sheng LIU Ying LI He-cheng FU De-lai ZHAI Xiao-qiang CHONG Tie(Department of Urology, Second Affiliated Hospital of Xi＇an Jiaotong University, Xi＇an 710004, China)

机构地区：[1]西安交通大学第二附属医院泌尿外科,陕西西安710004

出　　处：《现代泌尿外科杂志》2017年第1期58-62,共5页Journal of Modern Urology

摘　　要：目的研究雷帕霉素(RAPA)对体外培养的大鼠尿道成纤维细胞增殖、Ⅰ、Ⅲ胶原分泌以及TGF-β1表达的影响;方法组织块法原代培养雄性SD大鼠尿道成纤维细胞;MTT法检测不同浓度RAPA溶液在不同时间对鼠尿道成纤维细胞增殖和活力的影响;羟脯氨酸比色法测定RAPA干预后成纤维细胞培养上清液胶原含量的变化;RT-PCR检测雷帕霉素对TGF-β1、Ⅰ、Ⅲ胶原mRNA表达的影响;结果随浓度增加和时间延长体外培养鼠尿道成纤维细胞的吸光度(A值)逐步降低,实验组与对照组间存在显著性差异(P<0.05);羟脯氨酸比色法、RT-PCR测定不同浓度(0、10、20、40、80、160ng/mL)RAPA干预前后的成纤维细胞培养上清液胶原含量表达逐渐减少,TGF-β1、Ⅰ、Ⅲ胶原基因表达降低,各组间均存在显著性差异(P<0.05);结论雷帕霉素可抑制大鼠尿道成纤维细胞的增殖、减少胶原生成以及降低细胞内TGF-β1、Ⅰ、Ⅲ胶原的基因表达,在预防、治疗尿道狭窄中的作用值得进一步研究。Objective To investigate the effects of rapamycin （RAPA） on the proliferation of rat urethral fibroblasts cultured in vitro and the expressions of collagen Ⅰ -α1, collagen Ⅲ-α1 and TGF-β1. Methods The urethral fibroblasts of male SD rats were cultivated with tissue inoculation, which were treated with different concentrations of RAPA at different times. Cell proliferation was evaluated with MTT assay. The productions of collagen Ⅰ and Ⅲ were assessed with hydroxyproline colorimetric assay. The mRNA expressions of TGF-β1, collagen Ⅰ and Ⅲ in the fibroblasts were determined with RT-PCR. Results Compared with that in the control group, the A value of fibroblasts in the experimental group decreased with increasing concentration and time, and there was significant difference between the experimental group and the control group （P〈0.05）. Collagen content and expressions of collagen Ⅰ , Ⅲ and TGF-β1 mRNA treated with different concentrations of RAPA （0 ng/ mL, 10 ng/mL, 20 ng/mL, 40 ng/mL, 80 ng/mL, 160 ng/mL） reduced compared with those of the control group, and there were significant differences between the experimental group and control group （P〈0,05）. Conclusiorl Rapamycin can significantly inhibit the proliferation and collagen production of rat urethral fibroblasts in vitro. Moreover, rapamycin could reduce mRNA expressions of TGF-β1, collagen Ⅰ and Ⅲ. Further studies are deserved to investigate the effects of rapamycin on the prevention and treatment of urethral stricture.

关 键 词：雷帕霉素 尿道狭窄 成纤维细胞 

分 类 号：R34[医药卫生—基础医学]