细菌脂多糖体外诱导子宫内膜细胞炎症模型的建立  

Establishment of Inflammatory Model of Human Endometrial Cells Induced by Lipopolysaccharide in Vitro

作　　者：丁青[1] 尤昭玲[1] 陈怀敏[2] 杨静[3] 

机构地区：[1]湖南中医药大学,湖南长沙410007 [2]湖南中医药大学第一附属医院,湖南长沙410007 [3]湖南中医药大学2003级研究生班,湖南长沙410007

出　　处：《湖南中医学院学报》2006年第4期8-10,共3页Journal of Hunan College of Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(30271633);湖南省社会发展重大项目(01SSY1002-4)

摘　　要：目的探讨体外建立人子宫内膜细胞炎症模型,为子宫内膜炎症、出血及修复机制研究提供一个标准模型。方法取年龄20～40岁门诊女性患者,3月内未服用激素类药物,无菌条件下刮取宫腔前后壁内膜组织体外培养,以大肠杆菌细菌脂多糖(LPS)诱导子宫内膜细胞炎症,以培养细胞形态学和传统炎症指标作为炎症发生和发展的判断标准,以免疫细胞化学(ICC)方法鉴定细胞类型,以不同浓度的LPS作用于培养细胞,收集0、24、48、72h4个时段培养上清液用(ELISA)测定肿瘤坏死因子-α(TNF-α)、白细胞介素-β(IL-β)的含量。结果ICC显示培养细胞分别表达间质细胞标记物(Vimtein)、上皮细胞标记物(CK)及血管内皮细胞标记物(VEGF),部分细胞出现Vimtein与CK联合表达,炎症模型组培养上清液中TNF-α、IL-1β的含量相比同期空白组随LPS作用发展而逐渐上升(P<0.01)。结论培养的子宫内膜细胞类型包含上皮性、间质性、血管内皮源性细胞和少数具有双向分化潜能之细胞,在100ngm/LLPS作用下,体外培养的子宫内膜细胞诱导炎症达最合适浓度,炎症因子TNF-α、IL-1β含量增加表明模型建立成功。Objective To establish infection-Lipopolysaccharide model of human endometrial cells in vitro for a standard to detect inflammation with morphologic and biochemical index and provide a controlled research model and a standard for the study of endometrial inflection, bleeding and renovation. Methods The cultured tissue was scraped from patients who were 20 to 40 years old and without taking hormone in 3 months under aseptic conditions. The cultured human endometrial ceils were identified with ICC and stimulated by different concentration LPS, the ELISA were used to detect 0 h, 24 h, 48 h and 72 h concentration of TNF-α and IL-1 in the cell supernatants. Result The results of ICC demonstrated that the cultured cells could expressed interstitial cell marker Vimtein, epithelium cell marker CK and endothelial cells marker VEGF respectively, and some cells could expressed partly both Vimtein and CK. The concentration of TNF-α.IL-1 increased in the infection model of endometrial cells compared to the blank ones （P〈0.01）. Conclusion The cultured cells contain epithelium, interstitial cell, endotheliocyte and so on. In the cultured human endometrium cells there appear inflammatory pathologic characteristic under LPS-stimulated of the concentration 100ng/ mL., and the typical inflammatory factors TNF-α and IL-1β which shows that the establishment of inflammatory model is successful.

关 键 词：细菌脂多糖 子宫内膜细胞 炎症 模型 

分 类 号：R711.32[医药卫生—妇产科学]