基于HIF1A/EZH2/ANTXR2通路探讨罗氏内异方治疗子宫内膜异位症大鼠的作用机制  

Exploring the Mechanism of LUO’s Neiyi Prescription for the Treatment of Endometriosis Based on the HIF1A/EZH2/ANTXR2 Pathway

作　　者：裴芳利[1] 贾金金 陈晓鑫[2] 林树红 林增娴 吴李征 曾诚[1] PEI Fangli;JIA Jinjin;CHEN Xiaoxin;LIN Shuhong;LIN Zengxian;WU Lizheng;ZENG Cheng(The First Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510405 Guangdong,China;Guangzhou University of Chinese Medicine,Guangzhou 510006 Guangdong,China)

机构地区：[1]广州中医药大学第一附属医院,广东广州510405 [2]广州中医药大学,广东广州510006

出　　处：《中药新药与临床药理》2024年第7期993-1000,共8页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金项目(82305288,82074481);国家中医药传承创新中心科研专项(2022QN22);邓高丕名中医传承工作室(粤中医办函[2019]5号)。

摘　　要：目的基于缺氧诱导因子1A(HIF1A)/zeste增强子同源物2(EZH2)/炭疽热毒素受体2(ANTXR2)信号通路探讨罗氏内异方(又名益母调经化瘀合剂)对子宫内膜异位症(EMs)大鼠在位子宫内膜增殖及血管生成的干预作用及机制。方法将SD大鼠随机分为伪手术组、模型组、达那唑组(4.2 g·kg^(-1))及罗氏内异方低、高剂量组(15.74、31.48 g·kg^(-1)),每组8只。采用自体子宫内膜移植联合多因素干预法复制气滞血瘀型EMs病证结合大鼠模型。灌胃给药,每天1次,连续28 d。采用HE染色法观察子宫内膜组织病理变化;免疫组化法检测子宫内膜组织中增殖细胞蛋白67(Ki67)和血小板内皮细胞黏附分子1(CD31)蛋白表达水平;qRT-PCR及Western Blot法检测子宫内膜组织中HIF1A、EZH2、ANTXR2 mRNA及蛋白表达水平;Western Blot法检测子宫内膜组织中Yes关联蛋白1(YAP1)、吞噬细胞糖蛋白1(CD44)、β-连环蛋白(β-catenin)蛋白表达水平。结果与伪手术组相比,模型组大鼠在位子宫内膜组织上皮细胞增厚,间质细胞排列紊乱,炎性细胞增多;子宫内膜组织中Ki67、CD31的表达水平均显著升高(P<0.01),HIF1A、ANTXR2 mRNA及蛋白表达水平均显著升高(P<0.01),而EZH2 mRNA及蛋白表达水平显著下降(P<0.01),YAP1、CD44及β-catenin蛋白表达水平均显著升高(P<0.01)。与模型组比较,各给药组大鼠在位子宫内膜组织上皮层变薄,间质部细胞紊乱和炎症浸润均有改善,子宫内膜组织中Ki67、CD31的表达水平均显著降低(P<0.01);达那唑组与罗氏内异方高剂量组大鼠子宫内膜组织中HIF1A、ANTXR2 mRNA及蛋白表达水平均明显降低(P<0.05,P<0.01),EZH2 mRNA及蛋白表达水平明显升高(P<0.05,P<0.01);各给药组大鼠子宫内膜组织中YAP1、CD44及β-catenin蛋白表达水平均显著降低(P<0.05,P<0.01)。结论罗氏内异方可通过抑制在位子宫内膜细胞的增殖及血管生成能力来发挥治疗EMs的作用,其机制可能与抑制HIF1A/EZH2/Objective To investigate the intervention effect and mechanism of LUO’s Neiyi Prescription on the proliferation and angiogenesis of eutopic endometrium in rats with endometriosis(EMs)based on hypoxia-inducible factor 1A(HIF1A)/enhancer of zeste homolog 2(EZH2)/anthrax toxin receptor 2(ANTXR2)signaling pathway.Methods SD rats were randomly divided into sham operation group,model group,Danazol group(4.2 g·kg^(-1)),low-dose LUO’s Neiyi Prescription group and high-dose LUO’s Neiyi Prescription group(15.74,31.48 g·kg^(-1)),eight rats in each group.The rat model of EMs with qi stagnation and blood stasis syndrome was constructed by autologous endometrial transplantation combined with multi-factor intervention.Intragastric administration was given once a day for 28 consecutive days.The pathological changes of endometrial tissue were observed by HE staining.The expression levels of proliferating cell protein 67(Ki67)and platelet endothelial cell adhesion molecule 1(CD31)in endometrial tissue were detected by immunohistochemistry.The mRNA and protein expression levels of HIF1A,EZH2 and ANTXR2 in endometrial tissues were detected by qRT-PCR and Western Blot.The expression levels of YAP1,CD44 andβ-catenin in endometrial tissues were detected by Western Blot.Results Compared with the sham operation group,the epithelial cells of the eutopic endometrium of the model group were thickened,the interstitial cells were arranged disorderly,and the inflammatory cells increased.The expression levels of Ki67 and CD31 in endometrial tissues were significantly increased(P<0.01),the mRNA and protein expression levels of HIF1A and ANTXR2 were significantly increased(P<0.01),while the mRNA and protein expression levels of EZH2 were significantly decreased(P<0.01),and the protein expression levels of YAP1,CD44 andβ-catenin were significantly increased(P<0.01).Compared with the model group,the epithelial layer of the eutopic endometrial tissue of the rats in each administration group became thinner,the interstitial disorder and infl

关 键 词：子宫内膜异位症 罗氏内异方 益母调经化瘀合剂 在位子宫内膜 细胞增殖 血管生成 HIF1A/EZH2/ANTXR2信号通路 大鼠 

分 类 号：R285.5[医药卫生—中药学]