间苯三酚对泌尿平滑肌痉挛的调控作用机制  

The regulation mechanism of phloroglucinol in urinary smooth muscle spasm

作　　者：李聪睿 杜婷 燕红宇 罗明星 魏强[3] 苟马玲[1] LI Cong-rui;DU Ting;YAN Hong-yu;LUO Ming-xing;WEI Qiang;GOU Ma-ling(State Key Laboratory of Biotherapy,West China Hospital,Sichuan University,Chengdu 610041,China;Non-coding RNA and Drug Discovery Key Laboratory of Sichuan Province,Chengdu Medical College,Chengdu 610500,China;Dept of Urology,West China Hospital,Sichuan University,Chengdu 610041,China)

机构地区：[1]四川大学华西医院生物治疗国家重点实验室,四川成都610041 [2]成都医学院非编码RNA与药物四川省高校重点实验室,四川成都610500 [3]四川大学华西医院泌尿外科,四川成都610041

出　　处：《中国药理学通报》2023年第3期497-502,共6页Chinese Pharmacological Bulletin

基　　金：四川省科技计划项目(No 2022YFQ0004)。

摘　　要：目的探讨间苯三酚对膀胱平滑肌痉挛的调控作用及其机制。方法体外水平上,采用膀胱肌条实验验证间苯三酚对不同药物刺激下膀胱痉挛的缓解作用;同时,RT-qPCR和Western blot检测间苯三酚解痉挛作用对膀胱组织中钙离子信号通路相关基因和蛋白的表达水平。转录组学分析间苯三酚在解除膀胱痉挛时对其他相关通路基因的影响,用RT-qPCR来验证相关基因的表达情况。结果间苯三酚能够缓解不同药物刺激引起的膀胱痉挛,且解痉作用随浓度增高而增强,组织细胞中Calponin 1和MYLK3的表达水平升高,PKCα表达水平降低。RT-qPCR检测结果显示间苯三酚作用时,小鼠膀胱组织中Gprc5bG、Ppp2r5a、Chpt1、Prkar2b、Abcd2、Rasd1基因的表达水平均明显降低,与RNA-seq的测序结果一致。结论间苯三酚具有缓解膀胱平滑肌痉挛的作用,其作用机制与钙离子信号通路相关。间苯三酚作用于膀胱平滑肌后,抑制了细胞外Ca^(2+)内流,结合于细胞中的钙调蛋白4Ca^(2+)CaM复合物减少,导致游离的Calponin 1含量升高。钙调蛋白使得MLCK活化程度下降,MLC磷酸化减少,抑制了Mg^(2+)-ATP酶的作用,肌球蛋白横桥与细肌丝肌动蛋白分离,肌肉舒张。同时,间苯三酚还抑制Rasd1基因的表达,提示其在作用于膀胱平滑肌时还可能与细胞周期、蛋白磷酸化、胆碱代谢、ATP合成以及肿瘤信号通路相关。Aim To study the regulation and mechanism of phloroglucinol in bladder smooth muscle spasm.Methods In vitro the experiment used bladder muscle strip to verify the relieving effect of phloroglucinol on bladder spasm by different drugs.At the same time,RT-qPCR and Western blot were used to detect the expression levels of genes involved in the calcium signaling pathway caused by the antispasmodic effect of phloroglucinol.Results Phloroglucinol could relieve bladder spasm,and the antispasmodic effect was enhanced with the increase of concentration,and the expression of calponin 1 and MYLK3 in tissue cells increased.The results of RT-qPCR showed that the expression of Gprc5b G,Ppp2r5a,Chpt1,Prkar2b,Abcd2 and Rasd1 genes in mouse bladder tissue significantly decreased,which was consistent with the sequencing results of RNA-seq.Conclusions Phloroglucinol can relieve bladder smooth muscle spasm,and its mechanism is related to calcium signaling pathway.Meanwhile,phloroglucinol also inhibits the expression of Rasd1 gene,suggesting that it may be related to cell cycle,protein phosphorylation,choline metabolism,ATP synthesis and tumor-related pathways.

关 键 词：间苯三酚 机制研究 膀胱平滑肌 解痉作用 钙离子信号通路 转录组学 

分 类 号：R332[医药卫生—人体生理学] R322.62[医药卫生—基础医学] R322.74R348.1R394.2