摘要
Objective: The purpose of this study was to determine the role of Ureaplasma urealyticum-derived lipidassociated membrane proteins (LAMPs) in the host innate immune system, specifically their effect on Toll-like receptors (TLRs). Methods: LAMPs were derived from U. urea/yticum strains, and human amniotic epithelial cells (HAECs) were isolated from healthy full-term placentas. Cytokine concentrations were determined by enzyme-linked immunosorbent assay (ELISA) and TLR2 mRNA by real-time PCR. Expression of TLR2 was confirmed by Western blotting and immunohistochemistry. Results: LAMPs induced HAECs to produce inflammatory cytokines interleukin (IL)-6, IL-8, and tumor necrosis factor (TNF)-α. Cytokine production was reduced after blocking TLR2 using TLR2 inhibitor (anti-hTLR2-IgA). Conclusions: LAMPs isolated from U. urealyticum induced TLR2-dependent up-regulation of inflammatory genes and cytokines in HAECs.
目的:探讨解脲脲原体(Ureaplasma urealyticum)及其脂质相关膜蛋白(LAMPs)作用于人羊膜上皮细胞(HAECs)过程中白介素6(IL-6)、IL-8和肿瘤坏死因子α(TNF-α)的变化情况,阐明Toll样受体2(TLR2)的调控机制,明确解脲脲原体潜在的致病性。创新点:从解脲脲原体诱导炎症反应的分子机制入手,提出TLR2信号通路在其中的关键作用。方法:经TX-114处理萃取解脲脲原体获得LAMPs,将LAMPs和解脲脲原体分别感染HAECs,用酶联免疫吸附试验(ELISA)测定炎症细胞因子(IL-6、IL-8和TNF-α);采用实时聚合酶链反应(real-time PCR)测定TLR2 mRNA水平,用蛋白质印迹(Western blot)检测TLR2的表达量;经TLR2阻断剂(anti-h TLR2-IgA)处理后,测定相应炎症细胞因子。结论:解脲脲原体LAMPs能诱导HAECs的TLR2表达上调和炎症因子增加,从而发生炎症反应;TLR2受阻断后,炎症因子表达减少,炎症水平下降。TLR2在解脲脲原体LAMPs感染HAECs过程起关键作用。
基金
Project supported by the Zhejiang Provincial Natural Science Foundation of China(Nos.LY18H040001 and LY16H040003)
the Department of Education of Zhejiang Province(No.Y201534723)
the Science and Technology Development Project in Hangzhou(No.20160533B13),China