高迁移率族蛋白1启动Toll样受体介导的免疫炎症在OSAHS靶器官损害中的关键作用

Insufficient treatment impairs the effective control of OSAHS associated chronic diseases. It is very important for the organ protection of OSAHS to looking for the key factors as intervention targets. Our previous work showed the expression of Toll-like receptors (TLRs) after sleep increased more significantly than that before sleep, which could be inhibited by statins. So our hypothesis is high mobility group box protein 1 (HMGB1) starts and maintains the inflammation and injury in OSAHS by signaling through the partner molecule receptor - TLRs, which can be inhibited by statins. There will be three levels' study among human, mice and cells in this project. We are going to design the statins group and noninvasive positive pressure ventilation treatment group in the OSAHS patients, assess the expression of HMGB1, TLR2, TLR4, NF-κB and their downstream cytokines in plasma and white blood cells, and disclose its relationship with OSAHS prognosis. In the hypoxia model of mice , we are going to design the TLRs gene knockout groups, HMGB1 inhibitor groups and statins groups, and study the causation among the activation of HMGB1/TLRs/NF-κB, and inflammation injury in circulation, tissues and cells. The purpose of this project is to do the preliminary study to disclose the key role of HMGB1/TLRs /NF-κB as a upper stream signaling pathway in the circulation inflammation and organ injury in OSAHS, and to looking for the theoretical evidences for future new therapy target of OSAHS.

治疗不充分使OSAHS相关慢性疾病控制不佳，寻找其炎症损伤的核心分子机制作为靶点进行脏器保护十分重要。我们前期工作发现患者睡后toll样受体（TLRs）较睡前显著升高并可被他汀类药物抑制，故我们假设高迁移率族蛋白1（HMGB1）启动并维持了TLRs介导的OSAHS炎症损伤并可被他汀抑制。本研究设置OSAHS患者他汀类和正压通气治疗组，测定血浆和白细胞HMGB1、TLR2、TLR4、NF-κB及下游炎症因子表达，并进行长期随访；在动物缺氧模型中进行TLRs基因敲除、HMGB1抑制物和他汀干预机制研究，阐明HMGB1/TLRs/NF-κB在循环和组织的表达活化与炎症损伤的因果关系。本研究旨在为阐明HMGB1/TLRs /NF-κB在OSAHS循环炎症和器官损伤中的上游核心地位进行前期研究，为OSAHS器官保护寻找新的治疗靶点和理论依据。

.本研究按计划主要进行了以下2方面研究：.1. 阻塞性睡眠呼吸暂停低通气综合征（OSAHS）患者外周血循环中HMGB1/TLRs与循环炎症的关系：研究对象入组、标本及信息采集、测定血浆中可溶性HMGB1、TLR4、TLR2及TNFα水平（ELISA法），检测单核细胞和中性粒细胞中TLR4、TLR2表达情况（流式细胞仪）。.2. 完成大鼠低氧模型制备和标本留取，拟探索OSAHS独特病理生理过程中HMGB1/TLRs/NF-κB途径的表达与其在循环炎症和局部损伤中的因果关系。..主要研究成果：.1. OSAHS患者循环HMGB1在睡眠前后无显著性差异，但其配体TLR2和TLR4在基线水平（睡前）和睡眠前后却有显著性差异，而NPPV治疗可以消除这种差异，提示HMGB1的配体TLRs可能是OSAHS循环炎症的关键环节，也是未来干预的靶点之一。.2. 外周血中性粒细胞的TLR2和TLR4可能参与OSAHS的急性炎症过程，单核细胞的TLR2和TLR4可能参与OSAHS的慢性炎症反应，其中TLR4可能也参与了急性炎症过程； .3. OSAHS患者HMGB1/ TLR2/TLR4/TNFα和表达水平改变可能与夜间缺氧负荷（主要是时间负荷）以及微觉醒关系更为密切。.4. 睡前服用他汀类能够使OSAHS患者睡后HMGB1有较大幅度下降，显著消除OSAHS睡眠后外周血单核细胞TLR4的升高，提示他汀类可能具有潜在的抑制OSAHS炎症的作用，并有望成为OSAHS脏器保护综合治疗手段之一，但可能需要以达到稳态血药浓度为前提。