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
吴李征 1陈晓鑫 1孙博文 1裴芳利 2蓝丹彤 1邝梓君 3张诗媛 1曾诚2
作者信息
- 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
- 折叠
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(CoCl2)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(E2)、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,50 μmol·L-1 CoCi2处理在位ESCs 24 h可促进细胞增殖,显著增加HIF-1α mRNA和SF-1蛋白表达,而敲低HIF-1α可显著减少SF-1和E2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,E2、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.
Abstract
This study investigated the effect of Luoshi Neiyi Formula(LSNYF)on hypoxia-inducible factor-1α(HIF-1α)and ste-roidogenic factor 1(SF-1)in endometriosis(EMs),aiming to explore the mechanism of Luoshi Neiyi Formula in treating EMs.Immu-nohistochemistry and quantitative real-time PCR(qPCR)were employed to determine the expression of HIF-1α and SF-1 in the endo-metriotic tissue.Human primary endometrial stromal cells(ESCs)were extracted and identified,in which the expression levels of HIF-1α and SF-1 were measured by immunofluorescence and qPCR.Cobalt chloride(CoCl2)was used to simulate a low-oxygen envi-ronment in vitro,and the lentivirus with down-regulated expression of HIF-1α was constructed.The expression of HIF-1α or SF-1 was then determined by qPCR or Western blot.The effects of Luoshi Neiyi Formula-containing serum on the proliferation,migration,estra-diol(E2),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-1α and 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-1α and SF-1.Compared with the control group,treating eutopic ESCs with 50 pimol·L-1 CoCl2 for 24 h promoted cell proliferation and up-regulated the mRNA level of HIF-1α and protein level of SF-1,while knocking down HIF-1α down-regulated the expression of SF-1 and E2.Compared with the model group,the Luoshi Neiyi Formula-containing serum inhibited the proliferation and migration of ESCs and down-regulated the expression of E2,HIF-1α,and SF-1.Similarly,the treatment with Luoshi Neiyi Formula attenuated the pathologi-cal injury of the endometriotic tissue and down-regulated the mRNA and protein levels of HIF-1α and SF-1 in the rat model of EMs.In summary,LSNYF may treat EMs by alleviating hypoxia and inhibiting the HIF-1α/SF-1 axis.
关键词
子宫内膜异位症/罗氏内异方/HIF-1α/SF-1/缺氧Key words
endometriosis/Luoshi Neiyi Formula/HIF-1α/SF-1/hypoxia引用本文复制引用
基金项目
国家自然科学基金项目(82074481)
国家自然科学基金项目(82305288)
国家自然科学基金项目(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)
出版年
2024
NETL
NSTL
万方数据