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目的探讨子宫内膜异位症(EMS)患者异位及在位子宫内膜基质细胞在白细胞介素-1β(IL-1β)诱导培养条件下RANTES mRNA和蛋白表达模式的变化情况。方法对2006年9月至2007年6月南京医科大学第一附属医院收治的11例EMS患者的异位、在位内膜基质细胞和同期5例正常内膜基质细胞行分离培养,分别加入5、10、20μg/L的IL-1β刺激培养异位内膜基质细胞,选择最佳浓度的IL-1β刺激培养异位、在位和正常内膜基质细胞,采用RT-PCR及ELISA法从转录及转录后水平观察趋化因子RANTES表达模式的变化情况。结果10μg/LIL-1β诱导异位内膜基质细胞分泌RANTES蛋白能力强于5μg/L组,稳定性优于20μg/L组。异位内膜基质细胞于IL-1β刺激12h开始RANTES mRNA表达较在位及正常内膜显著增强,在位内膜基质细胞自48h开始RANTES mRNA表达较正常内膜显著增强。异位内膜自刺激培养24h开始RANTES蛋白分泌量明显高于在位内膜和正常内膜,96h可高达(926.52±97.19)ng/L,在位内膜自刺激培养60h开始RANTES蛋白分泌量较正常内膜显著增加,正常内膜基质细胞RANTES分泌量维持于(39.19±14.29)~(56.88±17.28)ng/L。结论EMS患者的异位、在位子宫内膜与正常子宫内膜相比,其RANTES表达特性已经发生了改变;异位内膜基质细胞可能是EMS患者腹腔液中高浓度RANTES的来源之一。
Objective To investigate the expression of RANTES mRNA and protein in ectopic and eutopic endometrial stromal cells of patients with endometriosis (EMS) induced by interleukin-1β (IL-1β). Methods Eighteen patients with ectopic EMS who were treated in the First Affiliated Hospital of Nanjing Medical University from September 2006 to June 2007 were enrolled in this study. Ectopic endometrial stromal cells and normal endometrial stromal cells were isolated and cultured in vitro. , 10,20μg / L IL-1βstimulated cultured ectopic endometrial stromal cells, select the best concentration of IL-1βstimulate ectopic, in situ and normal intima stromal cells, RT-PCR and ELISA from transcription And post-transcriptional level of chemokine RANTES expression patterns observed changes. Results 10μg / L IL-1β induced ectopic endometrial stromal cells to express RANTES protein stronger than 5μg / L group, the stability was better than 20μg / L group. The expression of RANTES mRNA in ectopic stromal cells was significantly increased at 12h after stimulation with IL-1β, and the expression of RANTES mRNA in eutopic endometrial stromal cells was significantly increased compared with that of normal endometrium since 48h. The excretion of RANTES protein in eutopic endometrium was significantly higher than that in eutopic endometrium and normal endometrium at 96 hours (926.52 ± 97.19) ng / L after cultured for 24h, and the secretion of RANTES protein at 60h Compared with normal endometrium, RANTES secretion in normal endometrium was significantly increased from (39.19 ± 14.29) ~ (56.88 ± 17.28) ng / L. Conclusion The ectopic and eutopic endometrium of EMS patients have changed their expression characteristics of RANTES compared with normal endometrium. Ectopic stromal cells may be one of the sources of high concentration of RANTES in peritoneal fluid of EMS patients.