补肾活血方调控Wnt/β-catenin信号通路改善复发性流产小鼠子宫螺旋动脉重铸的机制  

Kidney-tonifying and Blood-activating Prescription Regulates Wnt/β-catenin Signaling Pathway to Improve Uterine Spiral Artery Remodeling in Mouse Model of Recurrent Miscarriage

作　　者：李久现[1] 刘玉楠[2] 孙烁 孙建华 李香玉 陈萍[1] LI Jiuxian;LIU Yunan;SUN Shuo;SUN Jianhua;LI Xiangyu;CHEN Ping(The First Clinical Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450000,China;The First Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450000,China)

机构地区：[1]河南中医药大学第一附属医院,郑州450000 [2]河南中医药大学第一临床医学院,郑州450000

出　　处：《中国实验方剂学杂志》2024年第3期37-44,共8页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：河南省高等学校重点科研项目(20A360003);河南省中医药科学研究专项(2022JDZX011);第七批全国老中医药专家学术经验继承项目(国中医药人教函[2022]76号);河南省第二批中医药青苗人才培养项目(豫卫中医函[2021]16号)。

摘　　要：目的:观察补肾活血方对复发性流产模型小鼠子宫蜕膜组织Wnt/β-连环蛋白(β-catenin)信号通路及子宫螺旋动脉重铸的影响,探讨其作用机制。方法:以CBA/J×BALB/c小鼠合笼构建正常妊娠小鼠模型,以CBA/J×DBA/2小鼠合笼构建复发性流产小鼠模型,将复发性流产模型小鼠随机分为模型组、地屈孕酮组、中药低剂量组及中药高剂量组,以正常妊娠小鼠作为空白组,每组10只,各组分别给予药物治疗14 d。疗程结束后观察各组小鼠胚胎吸收率,苏木素-伊红(HE)染色观察蜕膜组织病理形态、螺旋动脉生理性转变率,实时荧光定量聚合酶链式反应(Real-time PCR)、蛋白免疫印迹法(Western blot)分别检测基质金属蛋白酶(MMP)-2、MMP-9、血管内皮生长因子(VEGF)及Wnt/β-catenin信号通路的mRNA与蛋白表达水平。结果:与空白组比较,模型组胚胎吸收率明显升高(P<0.05);子宫螺旋动脉生理性转变率明显降低(P<0.05);子宫蜕膜组织的细胞水肿变性,排列紊乱;螺旋动脉重铸关键因子MMP-2、MMP-9、VEGF及Wnt/β-catenin信号通路相关因子Wnt2、β-catenin、细胞周期蛋白D1(Cyclin D1)、c-Myc的mRNA与蛋白表达明显降低(P<0.05)。与模型组比较,中药低剂量组、中药高剂量组可以明显降低胚胎吸收率(P<0.05);明显提高螺旋动脉生理性转变率(P<0.05);改善子宫蜕膜组织的细胞形态、增加蜕膜血管数量;明显上调MMP-2、MMP-9、VEGF及Wnt/β-catenin信号通路相关因子的mRNA与蛋白表达(P<0.05)。结论:复发性流产存在子宫螺旋动脉重铸障碍,补肾活血方可以通过激活Wnt/β-catenin信号通路,增加VEGF、MMP-2及MMP-9的表达,促进子宫螺旋动脉重铸,从而治疗复发性流产。Objective:To observe the effects of the kidney-tonifying and blood-activating prescription on the Wnt/β-catenin signaling pathway and uterine spiral artery remodeling in a mouse model of recurrent miscarriage and to explore its underlying mechanism.Method:A mouse model of normal pregnancy was established by mating CBA/J mice with BALB/c mice.A mouse model of recurrent miscarriage was established by mating CBA/J mice with DBA/2 mice.The modeled mice of recurrent miscarriage were randomized into model,dydrogesterone,and low-and high-dose Chinese medicine groups.The mice in normal pregnancy were used as the control group.Each group consisted of 10 mice,and the drug administration lasted for 14 days.After the treatment,the embryo absorption rate of each group was recorded.Hematoxylin-eosin(HE)staining was employed to observe the pathological morphology of the uterine decidua,and the physiological transformation rate of spiral arteries(SPA)was evaluated.Real-time polymerase chain reaction(Real-time PCR)and Western blot were performed to determine the mRNA and protein levels,respectively,of matrix metalloproteinases(MMP)-2,MMP-9,vascular endothelial growth factor(VEGF),and Wnt/β-catenin signaling pathway.Result:Compared with the control group,the model group presented increased embryo absorption rate(P<0.05),decreased physiological transformation rate of uterine SPA(P<0.05),cellular swelling,degeneration,and disordered arrangement in the uterine decidua tissue,and down-regulated mRNA and protein levels of key factors involved in SPA remodeling(MMP-2,MMP-9,VEGF)and the Wnt/β-catenin signaling pathway(Wnt2,β-catenin,Cyclin D1,c-Myc)(P<0.05).Compared with the model group,both the low-and high-dose Chinese medicine reduced embryo absorption rate(P<0.05),increased SPA physiological transformation rate(P<0.05),improved uterine decidua tissue morphology,and increased decidua vessel count.Furthermore,they up-regulated the mRNA and protein levels of MMP-2,MMP-9,VEGF,and proteins in the Wnt/β-catenin signaling pathway(P<0.0

关 键 词：复发性流产 补肾活血方 子宫螺旋动脉重铸 绒毛外滋养细胞 Wnt/β-连环蛋白(β-catenin)信号通路 

分 类 号：R284[医药卫生—中药学] R285[医药卫生—中医学] R289R287R22R2-031R33R24