Department of Anesthesiology, University of Texas Medical Branch, Galveston, Texas, United States of America.
PLoS One. 2013 Jun 14;8(6):e65994. doi: 10.1371/journal.pone.0065994. Print 2013.
High mobility group box 1 (HMGB1), a highly conserved, ubiquitous protein, is released into the circulation during sterile inflammation (e.g. arthritis, trauma) and circulatory shock. It participates in the pathogenesis of delayed inflammatory responses and organ dysfunction. While several molecules have been identified that modulate the release of HMGB1, less attention has been paid to identify pharmacological inhibitors of the downstream inflammatory processes elicited by HMGB1 (C23-C45 disulfide C106 thiol form). In the current study, a cell-based medium-throughput screening of a 5000+ compound focused library of clinical drugs and drug-like compounds was performed in murine RAW264.7 macrophages, in order to identify modulators of HMGB1-induced tumor-necrosis factor alpha (TNFα) production. Clinically used drugs that suppressed HMGB1-induced TNFα production included glucocorticoids, beta agonists, and the anti-HIV compound indinavir. A re-screen of the NIH clinical compound library identified beta-agonists and various intracellular cAMP enhancers as compounds that potentiate the inhibitory effect of glucocorticoids on HMGB1-induced TNFα production. The molecular pathways involved in this synergistic anti-inflammatory effect are related, at least in part, to inhibition of TNFα mRNA synthesis via a synergistic suppression of ERK/IκB activation. Inhibition of TNFα production by prednisolone+salbutamol pretreatment was also confirmed in vivo in mice subjected to HMGB1 injection; this effect was more pronounced than the effect of either of the agents administered separately. The current study unveils several drug-like modulators of HMGB1-mediated inflammatory responses and offers pharmacological directions for the therapeutic suppression of inflammatory responses in HMGB1-dependent diseases.
高迁移率族蛋白 B1（HMGB1）是一种高度保守、普遍存在的蛋白质，在无菌性炎症（如关节炎、创伤）和循环性休克期间会释放到循环中。它参与延迟炎症反应和器官功能障碍的发病机制。虽然已经确定了几种调节 HMGB1 释放的分子，但对于鉴定 HMGB1 引发的下游炎症过程的药理学抑制剂（C23-C45 二硫键 C106 硫醇形式）的关注较少。在当前的研究中，在小鼠 RAW264.7 巨噬细胞中进行了针对临床药物和类药物化合物 5000+化合物的基于细胞的高通量筛选，以鉴定 HMGB1 诱导的肿瘤坏死因子-α（TNFα）产生的调节剂。抑制 HMGB1 诱导的 TNFα 产生的临床使用药物包括糖皮质激素、β激动剂和抗 HIV 化合物茚地那韦。对 NIH 临床化合物库的重新筛选确定了β激动剂和各种细胞内 cAMP 增强剂作为增强糖皮质激素对 HMGB1 诱导的 TNFα 产生的抑制作用的化合物。这种协同抗炎作用涉及的分子途径至少部分与通过协同抑制 ERK/IκB 激活来抑制 TNFα mRNA 合成有关。在接受 HMGB1 注射的小鼠体内也证实了预先用泼尼松龙+沙丁胺醇预处理抑制 TNFα 产生的作用；这种作用比单独使用任何一种药物的作用都更明显。本研究揭示了几种 HMGB1 介导的炎症反应的类药物调节剂，并为 HMGB1 依赖性疾病中炎症反应的治疗性抑制提供了药理学方向。
