补肾助孕方调节线粒体功能对子宫内膜血管生成的影响  

Effects of Bushen Zhuyun Prescription on Endometrial Angiogenesis by Regulating Mitochondrial Function

作　　者：周玉玲 张静蕾 曹璨 孙莹 牛若冰 张英美 胡书婵 罗琳 贺明 ZHOU Yuling;ZHANG Jinglei;CAO Can;SUN Ying;NIU Ruobing;ZHANG Yingmei;HU Shuchan;LUO Lin;HE Ming(College(Institute)of Integrative Medicine,Hebei University of Chinese Medicine,Hebei Key Laboratory of Integrative Medicine on Liver-Kidney Patterns,Hebei Collaborative Innovation Center of Integrated Traditional and Western Medicine on Reproductive Disease,Shijiazhuang 050200,China)

机构地区：[1]河北中医药大学中西医结合学院(研究所),河北省中西医结合肝肾病证研究重点实验室,河北省中西医结合生殖疾病协同创新中心,河北石家庄050200

出　　处：《中国中医药信息杂志》2024年第1期91-96,共6页Chinese Journal of Information on Traditional Chinese Medicine

基　　金：国家自然科学基金(82074247);河北省自然科学基金(H2021423031);河北省研究生创新项目(CXZZSS2022103);河北省大学生创新创业训练计划项目(S202114432008);河北省大中学生科技创新能力培育专项(22E50130D、22E50135D)。

摘　　要：目的观察补肾助孕方调节线粒体功能对子宫内膜血管生成的影响,探讨其改善子宫内膜容受性的作用机制。方法建立控制性超促排卵(COH)小鼠模型,将小鼠随机分为正常组、模型组和补肾组,每组20只,补肾组予补肾助孕方灌胃11 d,正常组和模型组予生理盐水灌胃,统计小鼠胚胎种植数,HE染色观察小鼠子宫内膜形态,免疫荧光染色观察子宫内膜组织α-平滑肌肌动蛋白(α-SMA)表达。体外培养子宫内膜微血管内皮细胞(HEMECs),将细胞分为对照组、VEGFA组、补肾组和VEGFA+补肾组,分别予血管内皮生长因子A(VEGFA)和/或补肾助孕方含药血清干预,检测细胞线粒体呼吸链复合物Ⅰ~Ⅳ活性,ATP含量及增殖细胞核抗原(PCNA)、胱天蛋白酶(Caspase-3)蛋白表达。结果动物实验显示,与正常组比较,模型组小鼠胚胎种植数显著降低(P<0.05),子宫内膜组织α-SMA蛋白表达显著降低(P<0.05);与模型组比较,补肾组小鼠胚胎种植数显著增加(P<0.05),子宫内膜组织α-SMA蛋白表达显著升高(P<0.05)。细胞实验显示,补肾助孕方含药血清可以提高HEMECs线粒体呼吸链复合物Ⅰ~Ⅳ活性及ATP含量,促进PCNA蛋白表达、抑制Caspase-3蛋白表达(P<0.05,P<0.01)。结论补肾助孕方促进子宫内膜血管生成可能与改善线粒体功能有关。Objective To observe the effects of Bushen Zhuyun Prescription on regulating mitochondrial function and endometrial angiogenesis;To explore its mechanism of improving endometrial receptivity.Methods The mouse model of controlled ovarian hyperstimulation(COH)was established,and the mice were randomly divided into normal group,model group,and Bushen group,with 20 mice in each group.Bushen group received Bushen Zhuyun Prescription for gavage for 11 d,and the normal group and model group received normal saline for gavage.The number of embryo implantation was counted,the endometrial morphology was observed by HE staining,α-smooth muscle actin(α-SMA)expression was observed by immunofluorescence staining.Human endometrial microvascular endothelial cells(HEMECs)were cultured in vitro,they were divided into control group,VEGFA group,Bushen group and VEGFA+Bushen group,and were intervened with VEGFA and/or Bushen Zhuyun Prescription medicated serum.The activities of mitochondrial respiratory chain complexesⅠ–Ⅳ,the content of ATP,the expression of PCNA and Caspase-3 were detected.Results Animal experiment showed that,compared with the normal group,the number of embryo implantation in model group significantly decreased(P<0.05),α-SMA protein expression in endometrial tissue significantly decreased(P<0.05);compared with the model group,the number of embryo implantation in Bushen group significantly increased(P<0.05),α-SMA protein expression in endometrial tissue significantly increased(P<0.05).Cell experiment showed that,Bushen Zhuyun Prescription medicated serum could increase the activity of mitochondrial respiratory chain complexesⅠ–Ⅳand ATP content in HEMECs,promote PCNA protein expression,and inhibit Caspase-3 protein expression(P<0.05,P<0.01).Conclusion Bushen Zhuyun Prescription can promote endometrial angiogenesis through improving mitochondrial function.

关 键 词：补肾助孕方 肾主生殖 线粒体 血管生成 子宫内膜容受性 小鼠 

分 类 号：R285.5[医药卫生—中药学]