Reduced-HMGB1 suppresses poly(I:C)-induced inflammation in keratinocytes
Reduced-HMGB1 suppresses poly(I:C)-induced inflammation in keratinocytes
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DOI：
10.1016/j.jdermsci.2018.01.007
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发表时间：
2018-05-01
影响因子：
4.6
通讯作者：
Sayama, Koji
中科院分区：
文献类型：
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作者：
Mori, Hideki;Murakami, Masamoto;Sayama, Koji
背景：高迁移率族蛋白1（HMGB 1）是一种核蛋白，具有稳定DNA和促进基因转录的作用。此外，细胞应激或死亡诱导HMGBI释放到细胞膜外，其中HMGBI作为警报蛋白发挥功能，与其他细胞因子、损伤相关分子模式（DAMP）和病原体相关分子模式（PAMP）组合引起炎症反应。（以前称为趋化-HMGB 1）对正常人角质形成细胞（NHK）中聚肌苷-聚胞苷酸[poly（I：C）]诱导的炎症的影响。C）-Toll样受体3（TLR 3）、视黄酸诱导基因-I（RIG-I）和黑素瘤分化相关蛋白5（MDA 5）途径，包括I κ B α、核因子（NF）-κ B p65、促分裂原活化蛋白激酶（MAPK）和干扰素调节因子3（IRF 3），并评估这些通路是否参与HMGB 1对NHK中poly（I：C）诱导的炎症的抑制。进行免疫沉淀，以了解是否HMGBI可以结合到聚（I：C），并进行免疫荧光染色和流式细胞仪分析，以检查是否减少HMGB干扰细胞摄取的聚（I：C）易位（可能通过内吞作用）。此外，还原型HMGB 1，而不是二硫化物型HMGB 1，对poly（I：C）诱导的NHK炎症具有抑制作用，表明外源性HMGB 1氧化还原状态的重要性。用还原型HMGB 1预处理抑制了NHK中由poly（I：C）刺激诱导的I κ B α、NF-κ B p65和IRF 3的磷酸化;然而，p38、细胞外信号调节激酶（ERK）和c-Jun N末端激酶（JNK）的磷酸化不受影响。二硫化物-HMGB 1与聚（I：C）形成复合物，还原和氧化的HMGB 1也是如此，尽管程度较低。免疫荧光染色和流式细胞仪分析表明，还原型HMGB干扰了细胞对poly（I：C）易位的摄取（可能是通过内吞作用）。结论：还原型HMGB 1预处理可改善NHK中poly（I：C）介导的炎症反应。(C)2018日本皮肤病研究学会。Elsevier B. V.出版，保留所有权利。
Background: High mobility group box 1 (HMGB1) is a nuclear protein that stabilizes DNA and facilitates gene transcription. Additionally, cell stress or death induces the release of HMGBI outside the cell membrane, where HMGBI functions as an alarmin, causing an inflammatory response in combination with other cytokines, damage-associated molecular patterns (DAMPs), and pathogen-associated molecular patterns (PAMPs).Objective: To evaluate the effect of reduced-HMGB1 (previously termed chemoattractive-HMGB1) on polyinosine-polycytidylic acid [poly(I:C)]-induced inflammation in normal human keratinocytes (NHKs).Methods: We focused on downstream components of the poly(I:C)-Toll-like receptor 3 (TLR3), retinoic acid-inducible gene-I (RIG-I), and melanoma differentiation-associated protein 5 (MDA5) pathways, including I kappa B alpha, nuclear factor (NF)-kappa B p65, mitogen-activated protein kinase (MAPK), and interferon regulatory factor 3 (IRF3), and assessed whether these pathways are involved in the suppression of poly(I:C)-induced inflammation in NHKs by HMGB1. An immunoprecipitation was performed to know whether HMGBI could bind to poly(I:C), and immunofluorescence staining and flow cytometric analysis were performed to check whether reduced-HMGB interferes with cellular uptake of poly(I:C) translocation (possibly by endocytosis).Results: Application of exogenous HMGB1 before, but not after, exerted a suppressive effect on poly(I:C)induced inflammation in NHKs. In addition, reduced-HMGB1, but not disulfide-HMGB1, exerted a suppressive effect on poly(I:C)-induced inflammation in NHKs, suggesting the importance of the redox status of exogenous HMGB1. Pre-treatment with reduced-HMGB1 inhibited the phosphorylation of I kappa B alpha, NF-kappa B p65, and IRF3 induced by poly(I:C) stimulation in NHKs; however, phosphorylation of p38, extracellular signal-regulated kinase (ERK), and c-Jun N-terminal kinase (JNK) was unaffected. Disulfide-HMGB1 formed a complex with poly(I:C), as did reduced- and oxidized-HMGB1, albeit to a lesser extent. Immunofluorescence staining and flow cytometric analysis indicated that reduced-HMGB interferes with cellular uptake of poly(I:C) translocation (possibly by endocytosis).Conclusion: These findings suggest that pre-treatment with reduced-HMGB1 ameliorates poly(I:C)-mediated inflammation in NHKs. (C) 2018 Japanese Society for Investigative Dermatology. Published by Elsevier B.V. All rights reserved.