罗氏内异方调控缺氧干预子宫内膜异位症SF-1的机制研究  

Mechanism of Luoshi Neiyi Formula in treatment of SF-1 on endometriosis by improving hypoxia

作　　者：吴李征 陈晓鑫[1] 孙博文 裴芳利[2] 蓝丹彤 邝梓君 张诗媛 曾诚[2] WU Li-zheng;CHEN Xiao-xin;SUN Bo-wen;PEI Fang-li;LAN Dan-tong;KUANG Zi-jun;ZHANG Shi-yuan;ZENG Cheng(Guangzhou University of Chinese Medicine,Guangzhou 510006,China;the First Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510405,China;the First People's Hospitol of Foshan,Foshan 528000,China)

机构地区：[1]广州中医药大学,广东广州510006 [2]广州中医药大学第一附属医院,广东广州510405 [3]佛山市第一人民医院,广东佛山528000

出　　处：《中国中药杂志》2024年第18期4967-4976,共10页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(82074481,82305288,82374500);国家中医药传承创新中心科研专项(2022QN22);广州中医药大学第一附属医院青优人才项目(中医一院[2023]186号);邓高丕名中医传承工作室项目(粤中医办函[2019]5号)。

摘　　要：观察罗氏内异方对子宫内膜异位症(EMs)中缺氧诱导因子-1α(HIF-1α)和类固醇生成因子1(SF-1)的影响,探索罗氏内异方对EMs的治疗机制。免疫组化及qPCR法检测临床EMs组织中HIF-1α和SF-1的表达。提取鉴定人原代子宫内膜间质细胞(ESCs),免疫荧光及qPCR法检测HIF-1α和SF-1的表达。用氯化钴(CoCl_(2))模拟体外缺氧环境,构建HIF-1α敲低慢病毒,qPCR或蛋白免疫印迹检测HIF-1α或SF-1的表达。观察罗氏内异方含药血清对缺氧环境下在位ESCs增殖、迁移、雌激素(E_(2))、HIF-1α和SF-1的影响。采用自体子宫内膜移植法联合多因素干预法建立气滞血瘀证EMs大鼠模型,观察罗氏内异方对大鼠异位子宫内膜组织中HIF-1α和SF-1的影响。结果提示,与正常子宫内膜组织及间质细胞相比,异位内膜组织及间质细胞中HIF-1α和SF-1的蛋白和mRNA水平均升高。与对照组相比,50μmol·L^(-1)CoCl_(2)处理在位ESCs 24 h可促进细胞增殖,显著增加HIF-1αmRNA和SF-1蛋白表达,而敲低HIF-1α可显著减少SF-1和E_(2)表达。与模型组相比,罗氏内异方含药血清组在位ESCs的增殖及迁移能力下降,E_(2)、HIF-1α和SF-1表达下降。同样,EMs大鼠经罗氏内异方治疗后,异位内膜组织病理损伤减轻,HIF-1α、SF-1的表达水平也显著降低。综上,罗氏内异方对EMs的治疗作用可能与改善缺氧环境,抑制HIF-1α/SF-1轴有关。This study investigated the effect of Luoshi Neiyi Formula(LSNYF)on hypoxia-inducible factor-1α(HIF-1α)and steroidogenic factor 1(SF-1)in endometriosis(EMs),aiming to explore the mechanism of Luoshi Neiyi Formula in treating EMs.Immunohistochemistry and quantitative real-time PCR(qPCR)were employed to determine the expression of HIF-1αand SF-1 in the endometriotic tissue.Human primary endometrial stromal cells(ESCs)were extracted and identified,in which the expression levels of HIF-1αand SF-1 were measured by immunofluorescence and qPCR.Cobalt chloride(CoCl_(2))was used to simulate a low-oxygen environment in vitro,and the lentivirus with down-regulated expression of HIF-1αwas constructed.The expression of HIF-1αor SF-1 was then determined by qPCR or Western blot.The effects of Luoshi Neiyi Formula-containing serum on the proliferation,migration,estradiol(E_(2)),HIF-1α,and SF-1 of ESCs in the low-oxygen environment were observed.Finally,the rat model of EMs with the syndrome of Qi stagnation and blood stasis was established by multi-factor intervention combined with autologous endometrium transplantation,and the effect of Luoshi Neiyi Formula on HIF-1αand SF-1 in the endometriotic tissue were studied.Compared with the normal endo-metrial tissue and ESCs,the endometriotic tissue and ectopic ESCs presented up-regulated mRNA and protein levels of HIF-1αand SF-1.Compared with the control group,treating eutopic ESCs with 50μmol·L^(-1)CoCl_(2)for 24 h promoted cell proliferation and up-regulated the mRNA level of HIF-1αand protein level of SF-1,while knocking down HIF-1αdown-regulated the expression of SF-1 and E_(2).Compared with the model group,the Luoshi Neiyi Formula-containing serum inhibited the proliferation and migration of ESCs and down-regulated the expression of E_(2),HIF-1α,and SF-1.Similarly,the treatment with Luoshi Neiyi Formula attenuated the pathological injury of the endometriotic tissue and down-regulated the mRNA and protein levels of HIF-1αand SF-1 in the rat model of EMs.In summary,

关 键 词：子宫内膜异位症 罗氏内异方 HIF-1Α SF-1 缺氧 

分 类 号：R285[医药卫生—中药学]