机械敏感通道Piezo1在子痫前期患者胎盘绒毛膜板动脉血管中的表达及意义

Expression and significance of the mechanosensitive channel Piezo1 in human placental chorionic arteries of pre-eclampsia patients

目的探讨机械敏感离子通道Piezo1在子痫前期(pre-eclampsia,PE)患者胎盘绒毛膜板动脉(human placental chorionic arteries,HPCA)中的表达及其意义。方法收集西南医科大学附属医院产科剖宫产分娩的孕妇的HPCA组织,包括血压正常的对照(normal control,NC)组、子痫前期组及重度子痫前期(sever preeclampsia,SPE)组,每组各10例。采用免疫荧光染色法检测NC组、PE组、SPE组孕妇HPCA中Piezo1和内皮标志性分子CD31的表达情况。采用实时荧光定量PCR法(real-time quantitative PCR,qPCR)检测NC组、PE组、SPE组孕妇HPCA中Piezol的mRNA相对表达;蛋白免疫印记法(Western blotting,WB)检测NC组、PE组、SPE组孕妇HPCA中Piezol的蛋白相对表达。过氧化氢(H_(2)O_(2))模拟氧化应激作用于人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVEC),采用qPCR法检测NC组、H_(2)O_(2)处理6 h组、H_(2)O_(2)处理12 h组HUVEC细胞中Piezol的mRNA相对表达;采用WB法检测NC组、H_(2)O_(2)处理6 h组、H_(2)O_(2)处理12 h组HUVEC细胞中Piezol的蛋白相对表达。结果Piezo1与内皮标志性分子CD31在NC组、PE组和SPE组孕妇HPCA组织中均存在共定位。Piezol的mRNA表达在NC组、PE组和SPE组孕妇HPCA组织中呈下降趋势,差异有统计学意义(P<0.05)。Piezol的蛋白表达在NC组、PE组和SPE组孕妇HPCA组织中呈下降趋势,差异有统计学意义(P<0.05)。在对照组、H_(2)O_(2)处理6 h组和H_(2)O_(2)处理12 h组HUVEC细胞中Piezo1的mRNA表达呈下降趋势,差异有统计学意义(P<0.05)。在对照组、H_(2)O_(2)处理6 h组和H_(2)O_(2)处理12 h组HUVEC细胞中Piezo1的蛋白表达呈下降趋势,差异有统计学意义(P<0.05)。结论Piezo1表达于HPCA组织内皮细胞中。Piezo1的异常表达可能与子痫前期的病理生理改变有关。子痫前期发生发展过程中氧化应激反应可导致Piezol表达下降。

Objective To investigate the expression and significance of the mechanosensitive ion channel Piezo1 in human placental chorionic arteries(HPCA)of pre-eclampsia(PE)patients.Methods HPCA tissues were collected from pregnant women undergoing cesarean section delivery at the Department of Obstetrics,Affiliated Hospital of Southwest Medical University.The samples included a normal control(NC)group,a PE group,and a severe pre-eclampsia(SPE)group,each comprising 10 cases.Immunofluorescence staining was used to detect the expression of Piezo1 and the endothelial marker CD31 in the HPCA of the NC,PE,and SPE groups.Real-time quantitative PCR(qPCR)was employed to measure the relative mRNA expression of Piezo1 in the HPCA of the NC,PE,and SPE groups.Western blotting(WB)was conducted to assess the relative protein expression of Piezo1 in the HPCA of the NC,PE,and SPE groups.Hydrogen peroxide(H_(2)O_(2))was used to simulate oxidative stress in human umbilical vein endothelial cells(HUVECs),and qPCR and WB were performed to evaluate the mRNA and protein expression of Piezo1 in the NC group,H_(2)O_(2)-treated 6 h group,and H_(2)O_(2)-treated 12 h group of HUVECs.Results Piezo1 co-localized with the endothelial marker CD31 in the HPCA tissues of the NC,PE,and SPE groups.The mRNA expression of Piezo1 showed a decreasing trend in the HPCA tissues of the NC,PE,and SPE groups,with statistically significant differences(P<0.05).Similarly,the protein expression of Piezo1 exhibited a decreasing trend in the HPCA tissues of the NC,PE,and SPE groups,with statistically significant differences(P<0.05).In HUVECs,both mRNA and protein expression of Piezo1 showed a decreasing trend in the NC group,H_(2)O_(2)-treated 6 h group,and H_(2)O_(2)-treated 12 h group,with statistically significant differences(P<0.05).Conclusion Piezo1 is expressed in endothelial cells of HPCA tissues,and its abnormal expression may be associated with the pathophysiological changes in pre-eclampsia.The occurrence and development of pre-eclampsia-related oxidative stress response may lead to decreased expression of Piezo1.