Liu Jianing, Wang Cong, Wang Wenyu, Ding Ning, Liu Jiaxin, Liu Hanwen, Wen Jiliang, Sun Wendong, Zu Shulu, Zhang Xiulin, Yan Jieke
Department of Kidney Transplantation, Multidisciplinary Innovation Center for Nephrology, The Second Hospital of Shandong University, Jinan, Shandong, China.
Department of Urology, The Second Hospital of Shandong University, Jinan, Shandong, China.
Front Pharmacol. 2024 Jun 26;15:1410565. doi: 10.3389/fphar.2024.1410565. eCollection 2024.
We aimed to investigate the expression and motor modulatory roles of several mechano-sensitive channels (MSCs) in human ureter. Human proximal ureters were obtained from eighty patients subjected to nephrectomy. Expression of MSCs at mRNA, protein and functional levels were examined. Contractions of longitudinal ureter strips were recorded in organ bath. A fluorescent probe Diaminofluoresceins was used to measure nitric oxide (NO). RT-PCR analyses revealed predominant expression of Piezo1 and TRPV2 mRNA in intact ureter and mucosa. Immunofluorescence assays indicate proteins of MSCs (Piezo1/Piezo2, TRPV2 and TRPV4) were mainly distributed in the urothelium. Ca2+ imaging confirmed functional expression of TRPV2, TRPV4 and Piezo1 in cultured urothelial cells. Specific agonists of Piezo1 (Yoda1, 3-300 μM) and TRPV2 (cannabidiol, 3-300 μM) attenuated the frequency of ureteral contractions in a dose-dependent manner while the TRPV4 agonist GSK1016790A (100 nM-1 μM) exerted no effect. The inhibitory effects of Piezo1 and TRPV2 agonists were significantly blocked by the selective antagonists (Dooku 1 for Piezo1, Tranilast for TRPV2), removal of the mucosa, and pretreatment with NO synthase inhibitor L-NAME (10 μM). Yoda1 (30 μM) and cannabidiol (50 μM) increased production of NO in cultured urothelial cells. Our results suggest that activation of Piezo1 or TRPV2 evokes NO production and release from mucosa that may mediate mechanical stimulus-induced reduction of ureter contractions. Our findings support the idea that targeting Piezo1 and TRPV2 channels may be a promising pharmacological strategy for ureter stone passage or colic pain relief.
我们旨在研究几种机械敏感通道(MSCs)在人输尿管中的表达及其对运动的调节作用。从80例行肾切除术的患者中获取人近端输尿管。检测了MSCs在mRNA、蛋白质和功能水平的表达。在器官浴槽中记录输尿管纵行肌条的收缩情况。使用荧光探针二氨基荧光素测量一氧化氮(NO)。逆转录聚合酶链反应(RT-PCR)分析显示,Piezo1和TRPV2 mRNA在完整输尿管和黏膜中表达占主导。免疫荧光分析表明,MSCs(Piezo1/Piezo2、TRPV2和TRPV4)的蛋白主要分布在尿路上皮。钙离子成像证实了TRPV2、TRPV4和Piezo1在培养的尿路上皮细胞中的功能性表达。Piezo1的特异性激动剂(Yoda1,3 - 300 μM)和TRPV2的特异性激动剂(大麻二酚,3 - 300 μM)以剂量依赖性方式减弱输尿管收缩频率,而TRPV4激动剂GSK1016790A(100 nM - 1 μM)则无作用。Piezo1和TRPV2激动剂的抑制作用被选择性拮抗剂(Piezo1的Dooku 1、TRPV2的曲尼司特)、去除黏膜以及用一氧化氮合酶抑制剂L-NAME(10 μM)预处理显著阻断。Yoda1(30 μM)和大麻二酚(50 μM)增加了培养的尿路上皮细胞中NO的产生。我们的结果表明,激活Piezo1或TRPV2可引起NO从黏膜产生和释放,这可能介导机械刺激诱导的输尿管收缩减少。我们的研究结果支持这样一种观点,即靶向Piezo1和TRPV2通道可能是一种有前景的药物策略,用于输尿管结石排出或缓解绞痛疼痛。