| 蒋锦杏,张 慧,任莉莉,高 宇,李 宁.滋养细胞胞内双链DNA感受器通过MAPK/IκB信号介导炎性细胞因子分泌[J].,2020,(8):1446-1450 |
| 滋养细胞胞内双链DNA感受器通过MAPK/IκB信号介导炎性细胞因子分泌 |
| Trophoblastic Intracellular Double-stranded DNA Sensors Mediate Inflammatory Cytokines Secretion via MAPK/ I?资B Signaling Pathway |
| 投稿时间：2019-11-06 修订日期：2019-11-30 |
| DOI：10.13241/j.cnki.pmb.2020.08.010 |
| 中文关键词: 胞内双链DNA感受器 滋养细胞 MAPK/IκB信号通路 炎性细胞因子 |
| 英文关键词: Intracellular double-stranded DNA sensor Trophoblast cells MAPK/IκB signaling pathway Inflammatory cytokines |
| 基金项目:广东省自然科学基金项目（2017A030310646，2019A1515011068）；广东省科技计划项目（2016A020215207）；深圳市科技计划项目（JCYJ20180228164515747）；中国博士后基金项目（2018M643371） |
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| 中文摘要: |
| 摘要 目的：探究滋养细胞能否感受胞内双链DNA刺激及其对炎性因子分泌的影响，揭示胎盘的免疫识别和免疫屏障功能，探讨妊娠期感染所致不良妊娠结局的发病机制。方法：利用人工合成的双链DNA模拟物poly(dA:dT)转染人滋养细胞系HTR-8/SVneo，Real-Time PCR方法检测滋养细胞胞内双链DNA识别受体的表达水平；Western Blot检测滋养细胞MAPK和IκB信号的活化情况；ELISA检测HTR-8/SVneo细胞培养上清中IL-6、IL-8、MCP-1、CXCL10的分泌水平。结果：Real-Time PCR结果表明，HTR-8/SVneo能够感受胞内双链DNA刺激并上调包括IFI16、AIM2、DHX36、DHX9、LRRFIP1、KU70、ZBP1/DAI和DDX41在内的多种双链DNA感受器的mRNA表达水平；Western Blot实验结果表明，滋养细胞识别双链DNA后能够促进MAPK和IκB信号通路活化，转染90分钟后ERK、JNK、p38和IκBα的磷酸化水平最高，其后随着时间逐渐减弱；加入IκB、p38和JNK特异性抑制剂能够抑制poly(dA:dT)介导的IL-8、IL-6和CXCL10分泌，但其分泌不受ERK抑制剂的影响；MCP-1的分泌能够被p38和JNK抑制剂阻断，但p38和ERK抑制剂不影响其分泌。结论：人滋养细胞存在功能性胞内双链DNA识别机制，活化的DNA识别信号能够激活MAPK和IκB信号通路，并通过IκB、p38或JNK信号介导IL-8、IL-6、MCP-1及CXCL10等细胞因子和趋化因子的分泌。 |
| 英文摘要: |
| ABSTRACT Objective: The study was preformed to investigate the expression pattern of intracellular double-stranded DNA sensors in human trophoblast cells and its effect on the secretion of inflammatory factors, to reveal the immune recognition and immune barrier function of placenta, and to better understand the pathogenesis of adverse pregnancy outcomes caused by pregnancy infection. Methods: Synthetic double-stranded DNA (dsDNA) mimics, poly(dA:dT), were transfected into human trophoblast cell line HTR-8/SVneo, double-stranded DNA sensors' mRNA were detected by quantitative real-time PCR and the phosphorylation levels of MAPK and IκB signaling were detected by Western Blot. ELISA was used to detect IL-6, IL-8, MCP-1 and CXCL10 secretion. Results: Real-Time PCR results showed that IFI16, AIM2, DHX36, DHX9, LRRFIP1, KU70, ZBP1/DAI and DDX41 mRNA levels were dramatically upregulated in HTR-8/SVneo cells. The phosphorylation levels of ERK, JNK, p38 and IκB were elevated at 90 minutes after poly(dA:dT) transfection, and then gradually decreased. The induction of IL-8, IL-6 and CXCL10 secretion is dependent on IκB, p38 and JNK signaling pathways; MCP-1 secretion is dependent on p38 and JNK signaling pathways. However, these cytokines and chemokines were not affected by ERK inhibitor. Conclusion: Human placental trophoblast cells express functional intracellular double-stranded DNA sensors, which can recognize double-stranded DNA and activate MAPK/IκB signaling pathways that mediate the secretion of IL-8, IL-6, MCP-1 and CXCL10. |
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