摘要
目的探讨机械感受器Yes相关蛋白1(YAP1)和压电型机械敏感离子通道蛋白2(PIEZO2)在小鼠膀胱尿路上皮细胞机械信号转导中的作用机制。方法使用FX-6000T细胞应力加载培养系统和YAP1特异性抑制剂维替泊芬(VP)处理小鼠膀胱尿路上皮细胞;通过实时荧光定量逆转录聚合酶链式反应(RT-qPCR)、蛋白质印迹法(WB)等实验检测PIEZO2、YAP1及其下游经典靶分子结缔组织生长因子(CTGF)等在mRNA及蛋白水平的表达情况,以及细胞ATP浓度变化。结果使用FX-6000T细胞应力加载培养系统拉伸刺激小鼠膀胱尿路上皮细胞后,荧光显微镜下观察到拉伸后的细胞直径长于拉伸前,差异有统计学意义(P<0.05)。拉伸组YAP1、PIEZO2及CTGF在mRNA和蛋白层面的表达水平均较未拉伸组高,差异有统计学意义(P<0.05)。VP处理有效降低了拉伸刺激后的膀胱尿路上皮细胞中YAP1、PIEZO2及CTGF在mRNA和蛋白层面的表达水平,差异有统计学意义(P<0.05)。拉伸刺激显著增加了细胞内的ATP浓度,而VP能够抑制ATP浓度增加,差异有统计学意义(P<0.0001)。结论拉伸刺激增加了膀胱尿路上皮细胞中YAP1和PIEZO2的表达,并促进了ATP的释放;VP则抑制了拉伸引起的YAP1活性增加和PIEZO2的高表达,进而减少了ATP的释放。小鼠膀胱尿路上皮细胞可能主要通过YAP1-PIEZO2-ATP途径来感知机械信号。
Objective To explore the roles of the mechanoreceptor Yes-associated protein 1(YAP1)and piezo type mechanosensitive ion channel component 2(PIEZO2)in mechanotransduction in mouse bladder urothelial cells.Methods Mouse bladder urothelial cells were subjected to mechanical stretching using the FX-6000T cell stretching system and treated with the YAP1-specific inhibitor verteporfin(VP).The expressions of PIEZO2,YAP1 and connective tissue growth factor(CTGF)at the mRNA and protein levels,as well as changes in cellular adenosine triphosphatase(ATP)concentration,were detected using reverse transcription quantitative PCR(RT-qPCR)and Western blotting(WB).Results After stretching stimulation,under the fluorescence microscope,it was observed that the diameter length of the stretched cells were longer than that before stretching,and the difference was statistically significant(P<0.05).The expressions of YAP1,PIEZO2 and CTGF at the mRNA and protein levels were increased in the stretched group compared to those of the non-stretched group(P<0.05).VP effectively reduced the expressions of YAP1,PIEZO2 and CTGF at the mRNA and protein levels after stretching stimulation(P<0.05).Stretching stimulation significantly increased the intracellular ATP concentration,while VP was able to inhibit the increase in ATP concentration,with a statistically significant difference(P<0.0001).Conclusion Stretching stimulation increased the expressions of YAP1 and PIEZO2 in bladder urothelial cells and promoted the release of ATP;verteporfin inhibited the increase in YAP1 activity and the overexpression of PIEZO2 caused by stretching,thereby reducing the release of ATP.It is suggested that mouse bladder urothelial cells may primarily sense mechanical signals through the YAP1-PIEZO2-ATP pathway.
作者
邵永祥
程猛
刘梦园
邢亮亮
范祖都
胡聪磊
幺立萍
张迁
刘飞
SHAO Yongxiang;CHENG Meng;LIU Mengyuan;XING Liangliang;FAN Zudu;HU Conglei;YAO Liping;ZHANG Qian;LIU Fei(Department of Urology,Xijing Hospital,Air Force Medical University,Xi'an 710032;Department of Urology,No.942 Hospital of Joint Logistics and Security Force of People's Liberation Army,Yinchuan 750003;Department of Air Service,Xijing Hospital,Air Force Medical University,Xi'an 710032;Graduate School of Xi'an Medical College,Xi'an 710021;Department of Digestive Medicine,Xijing Hospital,Air Force Medical University,Xi'an 710032,China)
出处
《现代泌尿外科杂志》
2025年第7期615-620,共6页
Journal of Modern Urology
基金
陕西省创新能力支撑计划项目(No.2024CX-GXPT-39)
国家科技重大专项研究项目(No.82070973)。
关键词
膀胱尿路上皮细胞
机械信号转导
压电型机械敏感离子通道蛋白
Yes相关蛋白
结缔组织生长因子
bladder urothelial cells
mechanical signal transduction
piezo type mechanosensitive ion channel component
Yes-associated protein 1
connective tissue growth factor
作者简介
通信作者:刘飞,副主任医师,副教授,硕士研究生导师。E-mail:liufeiyh@163.com;张迁,副主任医师,副教授,硕士研究生导师。E-mail:zhangqian2@fmmu.edu.cn;邵永祥,硕士研究生在读,主治医师。研究方向:尿控。E-mail:syx942yy@163.com。
