基于RhoA/Rho信号通路探讨产妇子宫平滑肌组织Thr38、KCa3.1、KLF4表达水平与产后出血的关系

The relationship between the expression levels of Thr38,KCa3.1 and KLF4 in uterine smooth muscle and postpartum hemorrhage based on RhoA/Rho signaling pathway

目的:基于RhoA/Rho信号通路探讨产妇子宫平滑肌组织CPI-17(Thr38)、KCa3.1、Krüppel样因子4(KLF4)表达水平与产后出血的关系。方法:选择80例宫缩乏力性产后出血产妇为观察组,选择同期80例无产后出血产妇为对照组。采用肌条等长张力测定法检测离体子宫平滑肌的自发收缩活动力,以及加入Rho激酶抑制剂(Y-27632)后的子宫平滑肌收缩活动力。采用实时荧光定量反转录聚合酶链反应法检测子宫平滑肌组织中RhoA、ROCKⅠ以及ROCKⅡmRNA表达水平,采用蛋白质印迹法检测RhoA、ROCKⅠ以及ROCKⅡ以及Thr38、KCa3.1、KLF4表达水平。采用Pearson相关分析RhoA蛋白以及子宫平滑肌收缩活动力与Thr38、KCa3.1、KLF4的关系。结果:观察组的子宫平滑肌收缩活动力、收缩频率以及收缩幅度均低于对照组(P<0.05~P<0.01),加入Y-27632后,2组子宫平滑肌收缩活动力均低于加入前(P<0.05)。观察组RhoA/Rho信号通路相关mRNA以及蛋白(RhoA、ROCKⅠ、ROCKⅡ)表达水平均低于对照组(P<0.01)。观察组Thr38、KCa3.1、KLF4表达水平均低于对照组(P<0.01)。Pearson相关分析显示,子宫平滑肌RhoA/Rho信号通路相关蛋白、收缩活动力与Thr38、KCa3.1、KLF4表达水平均呈正相关关系(P<0.05)。结论:宫缩乏力性产后出血产妇子宫平滑肌组织的RhoA/Rho信号通路以及Thr38、KCa3.1、KLF4表达下降,且与收缩能力降低呈正相关关系,共同参与产后出血的发生。

Objective:To investigate the relationship between the expression of p-cpi-17(Thr38),KCa3.1 and Kr in uterine smooth muscle tissue of puerpera and postpartum hemorrhage based on RhoA/Rho signaling pathway.Methods:A total of 80 patients of postpartum hemorrhage due to uterine inertia were selected as study group,and 80 patients without postpartum hemorrhage in the same period were selected as control group.The spontaneous contractile activity of isolated uterine smooth muscle and the contractile activity of uterine smooth muscle with Rho kinase inhibitor were measured by isometric tension test.Real-time fluorescence quantitative reverse transcription polymerase chain reaction method was used to detect the expression levels of RhoA,ROCKⅠ,and ROCKⅡmRNA in uterine smooth muscle tissues.Western blotting was used to detect the expression levels of RhoA,ROCKⅠ,ROCKⅡ,Thr38,KCa3.1,and KLF4.Pearson correlation was used to analyze the relationship between RhoA protein and uterine smooth muscle contractility and Thr38,KCa3.1,KLF4.Results:The uterine smooth muscle contraction activity,contraction frequency and contraction amplitude of the study group were lower than those of the control group(P<0.05 to P<0.01).After Y-27632 was added,the uterine smooth muscle contraction activity was lower than before(P<0.05).The expression levels of RhoA/Rho signaling pathway related mRNA and protein(RhoA,ROCKⅠ,ROCKⅡ)in the study group were lower than those in the control group(P<0.01).The expression levels of Thr38,KCa3.1,and KLF4 in the study group were lower than those in the control group(P<0.01).Pearson correlation analysis showed that RhoA/Rho signaling pathway related proteins and contractile activity were positively correlated with Thr38,KCa3.1,and KLF4 expression levels(P<0.05).Conclusions:The RhoA/Rho signaling pathway and expression of Thr38,KCa3.1,and KLF4 decrease in uterine smooth muscle tissue of parturients with postpartum hemorrhage due to uterine asthenia,and are positively correlated with the decrease of contractility,and jointly participate in the occurrence of postpartum hemorrhage.