重楼皂苷Ⅶ通过调控上皮间充质转化抑制子宫腺肌病异位内膜迁移和侵袭的分子机制  

Molecular mechanism of PolyphyllinⅦinhibiting ectopic endometrial migration and invasion of adenomyosis by regulating epithelial-mesenchymal transition

作　　者：石雅馨 王子璐 曹涵 高新宇 王领弟 刘志勇 师伟 SHI Yaxin;WANG Zilu;CAO Han;GAO Xinyu;WANG Lingdi;LIU Zhiyong;SHI Wei(First Clinical Medical College,Shandong University of Traditional Chinese Medicine,Jinan 250014,China;Affiliated Hospital of Shandong University of Traditional Chinese Medicine,Jinan 250014,China)

机构地区：[1]山东中医药大学第一临床医学院,济南250014 [2]山东中医药大学附属医院,济南250014

出　　处：《中华中医药杂志》2024年第5期2534-2541,共8页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金青年科学基金项目(No.82104916);山东省自然科学创新发展联合基金项目(No.ZR2023LZY024);山东省青年泰山学者项目(No.tsqn201909185);中国博士后科学基金(No.2022M722000)。

摘　　要：目的:探讨重楼皂苷Ⅶ(PPⅦ)通过调控上皮间充质转化(EMT)抑制子宫腺肌病(AM)异位内膜迁移和侵袭的作用机制。方法:通过生物信息学分析PPⅦ干预AM的关键靶点并通过体内外实验进行验证;他莫昔芬滴喂法建立AM小鼠模型并以PPⅦ进行干预,观察子宫形态学改变,苏木精-伊红(HE)染色观察子宫病理变化,免疫组化检测子宫钙黏蛋白1(CDH1)、钙黏蛋白2(CDH2)、基质金属蛋白酶2(MMP2)的表达;提取AM异位内膜细胞(AMDC),免疫荧光鉴定特征,CCK-8、Transwell实验检测PPⅦ对AMDC活性、迁移和侵袭能力的影响,RT-PCR和Western Blot检测CDH1、CDH2、MMP2的表达。结果:生信分析共获得381个PPⅦ靶点和1 630个AM差异表达基因,富集分析发现PPⅦ治疗AM主要涉及CDH1、CDH2、MMP2等关键靶点和EMT、迁移等重要生物学过程。体内实验显示与空白组比较,模型组子宫红肿、充血、增粗,子宫内膜-肌层结构紊乱,子宫内膜腺体侵入子宫肌层,子宫组织内CDH1表达显著降低(P<0.01),CDH2、MMP2表达显著升高(P<0.05,P<0.01);与模型组比较,米非司酮和PPⅦ组均可减少子宫内膜腺体侵入子宫肌层的数量,显著增加CDH1、降低CDH2、MMP2的表达水平(P<0.05)。体外实验证实,AMDC细胞具有间充质特性,PPⅦ均可抑制AMDC的迁移和侵袭能力,上调CDH1、下调CDH2、MMP2的mRNA和蛋白表达水平,呈剂量依赖效应。结论:PPⅦ可通过调控EMT进程抑制AM异位内膜的迁移和侵袭能力,为AM的防治与药物的研发提供新的探索方向与证据支持。Objective:To investigate the effect of polyphyllinⅦ(PPⅦ)on ectopic endometrial migration and invasion in adenomyosis(AM)by regulating epithelial mesenchymal transition(EMT).Methods:The key targets of PPⅦin the intervention of AM were analysed by bioinformatics and verified by in vivo and in vitro experiments.The AM mouse model was established by drip feeding of tamoxifen and treated with PPⅦ.The study observed morphological and pathological changes in the uterus using hematoxylin-eosin(HE)staining,and detected the expressions of cadherin 1(CDH1),cadherin 2(CDH2)and matrix metalloproteinase 2(MMP2)in the uterus through immunohistochemistry.Additionally,AMDC were isolated and subsequently characterized by immunofluorescence.The activity,migration and invasion of AMDC were evaluated using CCK-8 and Transwell experiments to detect the effect of PPⅦ.RT-PCR and Western Blot were used to detect the expression of CDH1,CDH2 and MMP2.Results:The bioinformatics analysis yielded 381 PPⅦtargets and 1630 differentially expressed genes in AM.Enrichment analysis revealed that PPⅦtreatment of AM primarily involved key targets,including CDH1,CDH2,and MMP2,and significant biological processes,such as EMT and migration.In the in vivo experiments,the uterus in the model group exhibited redness,congestion,and enlargement compared to the blank group.The structure of the endometrial-myometrium was disordered,with endometrial glands invading the myometrium,and the expression of CDH1 significantly decreased(P<0.01),CDH2 and MMP2expression significantly-increased(P<0.05,P<0.01),compared with the blank group.Compared with the model group,both the mifepristone and PPⅦgroups reduced the number of endometrial glands invading the myometrium,increased the expression levels of CDH1,decreased CDH2 and MMP2(P<0.05).In vitro experiments confirmed that AMDC exhibited mesenchymal properties.PPⅦinhibited the migration and invasive ability of AMDC,and had a dose-dependent effect on up-regulating the mRNA and protein expression levels o

关 键 词：子宫腺肌病 重楼皂苷Ⅶ 生物信息学 上皮间充质转化 迁移 侵袭 子宫腺肌病异位内膜细胞 

分 类 号：R285.5[医药卫生—中药学]