睾酮通过调控趋化因子受体CXCR3影响尿道狭窄疤痕成纤维细胞迁移的实验研究  

Experimental study on the changes of migration ability and the expression of chemokine receptor CXCR3 in human urethral stricture scar fibroblasts induced by testosterone

作　　者：王田龙[1] 薛竞东 俞仲伟[1] 冯超[2] 谢弘[2] 李锋[1] 章锋[1] 汪祖林[1] Wang Tianlong;Xue Jingdong;Yu Zhongwei;Feng Chao;Xie Hong;Li Feng;Zhang Feng;Wang Zulin(Department of Urology,Shanghai Eighth People's HospitalShanghai 200235;Department of Urology,Shanghai Jiaotong University affiliated Sixth People's HospitalShanghai 200233)

机构地区：[1]上海市第八人民医院泌尿外科,上海200235 [2]上海交通大学附属第六人民医院泌尿外科,上海200233

出　　处：《中国男科学杂志》2023年第1期30-35,47,共7页Chinese Journal of Andrology

基　　金：上海市卫生健康委员会科研课题No:20174Y0113。

摘　　要：目的探索人尿道狭窄疤痕成纤维细胞在不同双氢睾酮(DHT)环境中CXCR3表达水平变化及迁移变化、并探寻涉及的潜在相关生物信号通路。方法选取不同DHT浓度作用于人尿道狭窄疤痕成纤维细胞,Western-blotting检测不同组成纤维细胞CXCR3受体的表达情况。通过Transwell小室试验研究不同浓度的DHT对成纤维细胞迁移能力的影响。取用不同浓度DHT处理组和酒精刺激后的空白对照组,利用转录组测序技术(RNA-seq)分析寻找DHT与CXCR3潜在相关生物信号通路。结果Transwell实验证实,随着DHT浓度的逐步升高,成纤维细胞的迁移得以相应减弱。同时阶梯式升高浓度的DHT溶液环境可显著增加各组成纤维细胞CXCR3的表达,加入恩杂鲁胺拮抗雄激素受体(AR)后CXCR3受体表达明显下降。通过RNA测序后发现不同浓度雄激素处理后能显著改变部分基因的表达,主要富集于TGF-β信号通路、细胞黏合信号通路、类风湿炎症信号通路、MAPK信号通路、Ras信号通路、细胞运动骨架信号通路、Hippo信号通路、WNT信号通路等。结论在高雄激素环境中,人尿道狭窄疤痕成纤维细胞迁移有明显变化,其与CXCR3的表达成负相关,其潜在原因可能与TGF-β信号通路、细胞黏合信号通路等信号通路变化有关。Objective To explore the changes of migration ability and CXCR3 expression level of human urethral stricture scar fibroblasts in different dihydrotestosterone(DHT)environments and the possible underlying related biological signaling pathways.Methods Different concentrations of DHT were selected to act on human urethral stricture scar fibroblasts,and the effect of different concentrations of DHT on the migration ability of fibroblasts was studied by Transwell test.The expression of CXCR3 receptor of different fibroblasts was detected by Western blotting.The different concentrations of DHT group and the alcohol-stimulated blank control group were used for macroscopic analysis of transcriptome sequencing technology(RNA-seq)to find the potential related biological signaling pathways between DHT and CXCR3.Results Transwell experiments confirmed that with the gradual increase of DHT concentration,the migration ability of fibroblasts was correspondingly weakened.At the same time,the DHT solution environment with a stepwise increase in concentration can significantly increase the expression of CXCR3 in fibroblasts,and the expression of CXCR3 receptor was significantly decreased after adding enzalutamide to antagonize androgen receptor(AR).Androgen treatment can significantly change the expression of some genes,mainly enriched in TGF-βsignaling pathway,cell adhesion signaling pathway,rheumatoid inflammation signaling pathway,MAPK signaling pathway,Ras signaling pathway,cytoskeleton signaling pathway,Hippo signaling pathway,WNT signaling pathway,etc.Conclusion In the high concentrations of androgen environment,the migration of human urethral stricture scar fibroblasts has obvious changes,which is negatively related to the expression of CXCR3.The underlying reason may be related to TGF-βsignal pathway,cell adhesion signal pathway and other signal pathways.

关 键 词：睾酮 尿道狭窄 成纤维细胞 受体 CXCR3 

分 类 号：R695.4[医药卫生—泌尿科学]