Treg与促炎因子协同作用活化子宫内膜间质细胞Smad2信号通路  

Synergistic effect of Treg cells with proinflammatory cytokines on activating Smad2 signal pathway in endometrial stromal cells

作　　者：侯新新[1] 李大金[1] 王晓秋[1] 

机构地区：[1]复旦大学附属妇产科医院妇产科研究所,上海200011

出　　处：《现代免疫学》2016年第6期441-447,共7页Current Immunology

基　　金：国家自然科学基金青年科学基金(81200425)

摘　　要：本研究探讨了异位病灶调节性T细胞(Treg)与促炎细胞因子IL-1β、TNF-α协同作用对子宫内膜间质细胞Smad2信号通路的活化作用及其分子机制。采用原代分离培养子宫内膜异位症(简称内异症)患者在位及异位子宫内膜间质细胞(endometrial stromal cell,ESC);流式细胞术和ELISA法分别检测腹腔液中Treg的比例和TGF-β1的水平;免疫组织化学法分析正常、在位及异位内膜间质细胞中P-Smad2和Smad2的表达水平;细胞内Western法分析ESC Smad2信号通路的活化状况。结果显示:(1)与非内异症者相比,内异症患者盆腹腔内具有更多数量的Treg和更高水平的TGF-β1,且与疾病的进展呈正相关。(2)内异症患者在位及异位内膜具有更高的P-Smad2和Smad2水平。(3)异位病灶微环境中的Treg能够通过分泌TGF-β1与IL-1β或TNF-α发挥协同作用,可能通过激活ERK和p38信号通路活化ESC Smad2信号通路。ESC Smad2信号通路的活化可能通过多种分子机制影响ESC的生物学活性,从而导致疾病的发生和发展。This study investigated synergistic effect of regulatory T cells（Treg cells） and proinflammatory cytokines on activation of Smad2 signal pathway in endometrial stromal cell （ESC） and the underlying molecular mechanisms. ESC was separated from the eutopic endometrium of patients with endometriosis. Flow cytometry was performed to analyze the percentage of CD4＋ CD25＋ Foxp3＋ Treg cells in peritoneal fluid. The concentration of TGF-β1 in peritoneal fluid was quantified by ELISA kits. The expression of P-Smad2, Smad2 in the normal endometrium, eutopic endometrium and ectopic tissues were analyzed by immunohistochemistry. The activation of Smad2 signal pathway in ECS was detected using In-cell Western assay. Our result s showed that the percentage of CD4＋ CD25＋ Foxp3＋ Treg and the concentration of TGF-β1 were significantly increased in peritoneal fluid of women with endometriosis, compared with healthy fertile women, and the endometriosis patients at stage III-IV had the highest percentage of CD4＋ CD25＋ Foxp3＋ Treg and levels of TGF-β1. The eutopic endometrium and ectopic tissues had higher phosphorylation levels of Smad2, compared with that in the normal endometrium. The results also indicated that TGF-β1 secreted by Treg cells synergize with IL-β3 and TNF-α in the activation of Smad2 signal pathway through the activation of ERK and p38 signaling pathways in ESC, which may affect the biological activities of ESC that is conducive to the occurrence and development of the disease.

关 键 词：调节性T细胞 IL-1Β TNF—α 子宫内膜间质细胞 Smad2信号通路 

分 类 号：R711.71[医药卫生—妇产科学]