补肾安胎法调控Piezo1改善肾虚-黄体抑制病证结合流产大鼠母胎界面血管生成的机制  

Mechanism of regulating Piezo1 by tonifying kidney and calming fetus method to improve pregnancy outcomes of disease and syndrome combination rats of kidney deficiency and corpus luteum inhibition by promoting angiogenesis at the maternal-fetal interface

作　　者：谢宝珍 黄煦格 余兆吉 陈誉之 王然 曾粤睿 刘紫嫣 郜洁[2,3] 罗颂平[2,3] XIE Baozhen;HUANG Xuge;YU Zhaoji;CHEN Yuzhi;WANG Ran;ZENG Yuerui;LIU Ziyan;GAO Jie;LUO Songping(First School of Clinical Medicine,Guangzhou University of Chinese Medicine,Guangzhou 510405,China;Lingnan Medical Research Center of Guangzhou University of Chinese Medicine,Guangzhou 510405,China;First Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510405,China)

机构地区：[1]广州中医药大学第一临床医学院,广州510405 [2]广州中医药大学岭南医学研究中心,广州510405 [3]广州中医药大学第一附属医院,广州510405

出　　处：《中华中医药杂志》2023年第7期3065-3071,共7页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金面上项目(No.81873336);中医药传承与创新“百千万”人才工程(岐黄工程)——国家中医药领军人才支持计划(No.国中医人教发[2018]12号);广东省重点领域研发计划项目(No.2020B1111100003);市校(院)联合资助项目市重点实验室建设项目(No.202201020383)。

摘　　要：目的:阐明补肾安胎法介导Piezo1改善病证结合大鼠母胎界面血管重铸的作用机制。方法:构建肾虚-黄体抑制病证结合流产S D大鼠,将8~10周龄孕鼠随机分为6组,每组9只:空白对照(C o n t r o l)组、模型(Model)组(羟基脲45 mg/mL+米非司酮0.4 mg/mL)、模型+地屈孕酮(Progesterone)组(地屈孕酮0.31 mg/mL)、模型+减味寿胎丸(JSP)低剂量(JSP-L)组(JSP 82.23 mg/mL)、模型+减味寿胎丸中剂量(JSP-M)组(JSP 164.46 mg/mL)、模型+减味寿胎丸高剂量(JSP-H)组(JSP 328.92 mg/mL)。体视镜分析各组孕鼠胚胎及胚胎血管发育情况;小动物超声比较各组孕鼠子宫动脉血流阻力;小动物光声成像比较各组孕鼠胎盘血氧饱和度差异;苏木素-伊红(HE)染色观察各组孕鼠卵巢黄体数目、黄体血管及胎盘血管丰富度;免疫组织化学染色(IHC)和蛋白免疫印迹法(WB)检测各组孕鼠胎盘Piezo1及血管生成相关分子VEGFR2、VEGFR1及VEGFA的分布、表达及蛋白水平表达。结果:与Control组比较,Model组孕鼠子宫及胚胎湿重下降(P<0.05),胚胎直径缩小(P<0.05),流产率升高至65.73%(P<0.001);子宫动脉搏动指数PI及阻力指数RI升高(P<0.01);胎盘血氧饱和度下降(P<0.05);HE示妊娠黄体数目下降,妊娠黄体血管密度及胎盘血管丰富度不足;IHC示Model组孕鼠胎盘Piezo1及VEGFA表达降低(P<0.001),血管生成因子受体VEGFR2及VEGFR1表达升高(P<0.001);Western Blot检测示Model组孕鼠胎盘Piezo1表达下降(P<0.01),VEGFR2及VEGFR1表达升高(P<0.05)。与Model组比较,Progesterone组及JSP各剂量组流产率均下降(P<0.01);子宫动脉PI及RI下降(P<0.05);JSP-L及JSP-M组血氧饱和度升高(P<0.01);HE示Progesterone组及JSP各剂量组妊娠黄体及胎盘血管丰富度增加;IHC检测示JSP各剂量组孕鼠胎盘Piezo1及VEGFA表达升高(P<0.001);Western Blot检测示JSP-M组孕鼠胎盘Piezo1及VEGFR2表达增加(P<0.01)。结论:补肾安胎中药复方减味寿胎丸通过调控机械敏感性离子通�Objective:To elucidate the mechanism of how tonifying kidney and calming fetus method mediates Piezo1 to improve angiogenesis at the maternal-fetal interface in a disease and syndrome combination rat model.Methods:Kidney deficiency and corpus luteum inhibition rat abortion model was constructed.8-10 weeks old pregnant rats were randomly divided into 6 groups(9 rats in each group):blank control group,model group(hydroxyurea 45 mg/mL+mifepristone 0.4 mg/mL),model+dydrogesterone(progesterone 0.31 mg/mL),model+Jianwei Shoutai Pill(JSP)low dose group(JSP 82.23 mg/mL,JSP-L),model+JSP medium dose group(JSP 164.46 mg/mL,JSP-M),and model+JSP high dose group(JSP 328.92 mg/mL,JSP-H).The embryonic and its vascular development of each group was analyzed by somatoscopy.Small animal ultrasound was applied to analyze uterine artery flow resistance.Photoacoustic imaging was performed to analyze placental oxygen saturation.Hematoxylin-eosin(HE)staining was applied to analyze the number of ovarian corpus luteum,corpus luteum vessels and placental vascular abundance.Immunohistochemistry(IHC)was performed to analyze distribution and expression of Piezo1,VEGFR2 and VEGFR1,VEGFA in the placenta.Protein expression of Piezo,VEGFR2,VEGFR1 and VEGFA in the placenta were examined by Western Blot(WB).Results:Compared to the Control group,Model group showed a decrease in uterine and embryonic wet weight(P<0.05),a decrease in embryonic diameter(P<0.05)and an increase in abortion rate to 65.73%(P<0.001).It also showed an increase in uterine artery pulsatility index PI and resistance index RI(P<0.01),a decrease in placental oxygen saturation(P<0.05),a decrease in the number of corpus luteum and a decrease in placental vascular richness.IHC showed decreased expression of Piezo1 and VEGFA(P<0.001)and increased expression of VEGFR2 and VEGFR1 in Model group(P<0.001).WB also showed decreased expression of Piezo1(P<0.01)and increased expression of VEGFR2 and VEGFR1 in Model group(P<0.05).Compared to Model group,Progesterone group and the JSP groups s

关 键 词：补肾安胎法 减味寿胎丸 Piezo1 血管生成 自然流产 病证结合模型 

分 类 号：R285.5[医药卫生—中药学]