孕酮通过RhoA/ROCK信号通路促进子宫平滑肌收缩的机制研究  

Mechanism of progesterone promoting uterine smooth muscle contraction through RhoA/ROCK signaling pathway

作　　者：严小丽 陈诚 王丹 常青 YAN Xiaoli;CHEN Cheng;WANG Dan;CHANG Qing(Department of Gynecology and Obstetrics,First Affiliated Hospital,Army Medical University(Third Military Medical University),Chongqing,400038;Department of Gynecology and Obstetrics,Chongqing Hospital of University of Chinese Academy of Sciences(Chongqing General Hospital),Chongqing,401147,China)

机构地区：[1]陆军军医大学(第三军医大学)第一附属医院妇产科,重庆400038 [2]中国科学院大学重庆医院(重庆市人民医院)妇产科,重庆401147

出　　处：《陆军军医大学学报》2022年第12期1199-1206,共8页Journal of Army Medical University

基　　金：国家自然科学基金面上项目(81771619,81671494)。

摘　　要：目的 探讨孕酮通过RhoA/ROCK信号通路影响子宫平滑肌收缩的作用机制。方法 收集临产与未临产人子宫平滑肌组织各5例,采用Western blot和免疫组化检测肌球蛋白轻链20磷酸化(phosphorylation of myosin light chain 20,p-MLC20)、ROCK1、ROCK2的表达;分离培养子宫平滑肌细胞并经免疫荧光技术鉴定,激光共聚焦显微镜检测细胞内游离Ca;浓度([Ca;]i);孕酮受体拮抗剂米非司酮及孕酮(P4)干预子宫平滑肌细胞,Western blot检测细胞中p-MLC20及ROCK1、ROCK2的表达,以及ROCK抑制剂Y-27632与孕酮干预后子宫平滑肌细胞中p-MLC20的表达。结果 与未临产组织相比,临产子宫平滑肌组织中p-MLC20、ROCK1、ROCK2表达明显上升(P<0.05);孕酮诱导子宫平滑肌细胞后,细胞内ROCK1、ROCK2、p-MLC20水平增加(P<0.05);Y-27632与孕酮干预子宫平滑肌细胞后,细胞内p-MLC20水平下降(P<0.05)。结论 孕酮通过RhoA/ROCK信号通路调控p-MLC20水平,促进子宫平滑肌细胞收缩。Objective To investigate the mechanism of progesterone affecting the contractions of uterine smooth muscle through RhoA/ROCK signaling pathway. Methods The tissues of uterine smooth muscle were collected from 5 cases in labor and another 5 of non-labor, respectively. The expression levels of phosphorylation of myosin light chain(p-MLC20), ROCK1 and ROCK2 were detected by Western blotting and immunohistochemical(IHC) assay. Uterine smooth muscle cells were isolated and cultured, and then identified by immunofluorescence assay. Laser confocal microscopy was used to measure the concentration of intracellular free Ca;([Ca;]i) in the cells. In addition, the uterine smooth muscle cells were treated with progesterone+progesterone receptor antagonist, mifepristone, and with progesterone+ROCK inhibitor, Y-27632, respectively. Western blotting was performed to determine the expression of p-MLC20, ROCK1 and ROCK2 in the corresponding groups. Results As compared with the non-labor group, the expression of p-MLC20, ROCK1 and ROCK2 in the uterine smooth muscle tissues were significantly increased in the labor group(P<0.05). Progesterone induced the elevation of p-MLC20, ROCK1 and ROCK2 levels in the cultured uterine smooth muscle cells(P<0.05), while after the intervention of Y-27632+progesterone, the p-MLC20 level was greatly decreased(P<0.05). Conclusion Progesterone regulates p-MLC20 level through RhoA/ROCK signaling pathway and thus promotes the contractions of uterine smooth muscle cells.

关 键 词：孕酮 RhoA/ROCK信号通路 肌球蛋白轻链20磷酸化 子宫收缩 平滑肌细胞 

分 类 号：Q579.13[生物学—生物化学] R329.3[医药卫生—人体解剖和组织胚胎学] R714.3[医药卫生—基础医学]