孕激素对子宫内膜异位症患者在位内膜T淋巴细胞分泌的受激活调节因子表达的影响

Effects of Progesterone and Progestin on Expression of Regulated on Activation,Normal T Cell Expressed and Secreted in Eutopic Endometrium from Patients with Endometriosis

目的探讨正常T淋巴细胞表达和分泌的受激活调节因子(RANTES)与子宫内膜异位症(EM)发病的关系以及孕激素治疗的可行性。方法采集子宫内膜异位症患者术前以及放置左炔诺孕酮宫内节育系统(LNG-IUS)、口服甲羟孕酮(MPA)或注射促性腺激素释放激素类似物(GnRHa)后以及对照组的在位内膜,以半定量RT-PCR方法检测内膜组织中RANTES mRNA的表达。以100U/ml肿瘤坏死因子-α(TNFα)和不同浓度的孕酮(Po)与内膜细胞共孵育24h,观察Po对TNFα刺激RANTES分泌效应的影响。以不同浓度Po预处理细胞48h后,加入TNFα(100U/ml,16h),观察Po对TNFα刺激细胞分泌RANTES的抑制作用。采用ELISA方法检测细胞培养上清中RANTES的分泌量。结果药物干预前,EM组RANTES mRNA的相对表达量显著高于对照组(28.0±9.0vs.22.0±5.6,P<0.05)。放置LNG-IUS(24.0±4.2vs.25.9±4.2,P>0.05)或注射GnRHa(23.0±12.9vs.26.9±5.2,P>0.05)后,子宫内膜RANTES mRNA的表达差异无显著性;MPA组RANTES mRNA表达较用药前显著升高(42.6±3.1vs.24.3±5.7,P<0.05)。单纯Po刺激,培养子宫内膜细胞RANTES的分泌量无明显变化;同时以Po和TNFα刺激细胞,RANTES的分泌显著增加;Po预处理48h后,RANTES的分泌对TNFα的反应性显著减弱。结论EM患者在位内膜本身具有高趋化活性,孕激素防治EM有一定可行性。

Objective To investigate the effects of progesterone and progestin on the expressions of regulated on activation, normal T cell expressed and secreted （RANTES） in eutopic endometrium from patients with endometriosis. Methods We collected the samples of endometrium from patients with endometriosis before operation or after insertion of levenorgestrel releasing intrauterine system （LNG-IUS）, administration of oral medroxyprogesterone （ MPA）, or injection of gonadotrophic hormone releasing hormone agonist （GnRHa）. Reverse transcription-polymerase chain raction was used to assay the expression of RANTES mRNA. On the other hand, progesterone （Po） and tumor necrosis factor-α （TNFα） of different concentrations and different manners were used to treat cultured cells in vitro. RANTES secretion was evaluated in the culture medium using ELISA. In order to evaluate the effect of P0 on the secretion of RANTES under stimulation of TNFα, the cells were cultured in medium containing 100 U/ml TNFα and Po of different concentrations for 24 hours. After the pretreatment of Po for 48 hours at different concentrations, TNFα （ 100 U/ml, 16 h） was added to observe whether Po inhibits RANTES or not. Results The expression of RANTES mRNA in eutopic endometrium of patients with endometriosis was significantly higher than in control group （ 28.0 ± 9.0 vs. 22.0 ± 5.6, P 〈 0. 05 ）. Following the exposures to LNG-IUS （ 24.0 ± 4.2 vs. 25.9 ± 4.2, P 〉 0.05 ） or GnRHa （ 23.0 ± 12.9 vs. 26.9 ± 5.2, P 〉 0. 05 ）, the expression of RANTES mRNA had no change. MPA significantly increased the expression of RANTES mRNA （42. 6 ± 3.1 vs. 24.3 ± 5.7, P 〈 0.05 ）. Po itself had no significant effect on the secretion of RANTES. Stimulated by Po and TNFα at the same time, the secretion of RANTES significantly increased. After pretreatment with Po for 48 hours, the reaction of RANTES to the stimulating effect of TNFα was down-regulated. Conclusion The eutopic endometrium of patients with endometriosis has high chemotactic activity. It may be feasible to prevent and treat endometriosis with progestins.