中药血必净注射液调控血管内皮细胞TM表达抑制重症中暑DIC发生发展的机制研究

Severe heatstroke combined with DIC is a prominent pathological mechanism leading to the early death and developing into MODS. Once severe heatstroke combined with MODS the mortality is up to 30-60%. Previous studies have delineated that vascular endothelial injury is the key factor in the onset of DIC induced by severe heatstroke. Our previous clinical observations have found that Xuebijing significantly reduces the mortality of severe heatstroke patients. As for its mechanism, We have found that Xuebijing has protective effects on the multiple organs of severe heatstroke patients including the nervous system, circulation, blood clotting, liver, lung and intestines. Further studies confirmed that Xuebijing could inhibit severe heatstroke accompanied with DIC and rectify coagulation/anticoagulation imbalance, and its possible mechanism is closely related to the regulation of TM which means it plays an important role on increasing the expression of TM in endothelial cells and reducing the level of sTM scereted. However, the specific molecular mechanism of Xuebijing on regulating the expression of TM in severe heatstroke endothelial cells and decrease sTM has not been reported yet. Therefore, this study aims to elucidate the role and molecular mechanism of Xuebijing on regulating the expression of TM in severe heatstroke and inhibiting the occurrence and development of DIC through animals with heatstroke, endothelial cell model and clinical patients. This study will provide a theoretical basis for the clinical application of Xuebijing.

重症中暑合并MODS死亡率高达30-60%，合并DIC是导致重症中暑早期死亡和发展为MODS的重要因素。已有的研究表明，血管内皮损伤是启动重症中暑DIC的关键环节。课题组既往临床观察发现，血必净可显著降低重症中暑患者死亡率。对其机制研究发现，血必净可对重症中暑多脏器包括神经系统、循环、凝血、肝、肺、肠道等起到保护作用。并在进一步的研究中证实，血必净可以抑制重症中暑DIC、恢复凝血平衡，其可能的机制与调控TM密切相关，即通过上调内皮TM表达、降低sTM水平来发挥作用。然而，血必净如何调控重症中暑血管内皮细胞TM表达并降低sTM水平的具体分子机制尚未见报道。因此，本研究拟进一步以热打击动物、内皮细胞模型和临床患者为研究对象，全面分析血必净调控血管内皮TM表达从而抑制重症中暑DIC发生发展的作用和分子机制，为血必净的临床应用奠定理论基础。

重症中暑合并MODS死亡率高达30-60%，发生DIC是导致重症中暑早期死亡和发展为MODS的重要因素。研究表明，肠道菌群代谢产物TMAO可调控NETs表达，而NETs所致的免疫性血栓形成是DIC发生发展的关键环节。课题组既往临床观察发现，血必净可以抑制重症中暑DIC、恢复凝血平衡。进一步研究证实，血必净可以改善中暑大鼠肠道菌群结构，影响其代谢产物释放。预实验也发现，血必净可以降低重症中暑DIC患者血浆中肠道菌群代谢产物TMAO水平。然而，血必净如何通过调控肠道菌群代谢产物TAMO介导的NETs表达从而抑制重症中暑DIC发生发展的具体分子机制尚未明确。因此，本研究拟通过以热打击动物、中性粒细胞模型和临床中暑患者为研究对象，全面分析血必净调控肠道菌群代谢产物TAMO介导的NETs表达从而抑制重症中暑DIC发生发展的作用和分子机制，为血必净临床救治重症中暑奠定理论基础。我们在研究中发现：1.血必净能够改善重症中暑患者脏器功能及减轻全身炎症反应；2.中暑大鼠血清sTM及股动脉、股静脉中FPA表达增高；3.热打击对HUVECs TM及sTM表达的影响；4.血必净预处理能够改善中鼠大鼠凝血功能及减轻中暑大鼠血清中炎症因子水平；5.血必净可能通过NR4A2-KLF2途径改善凝血功能及减轻炎症反应。本项目对重症中暑、凝血功能紊乱、血必净治疗之间的分子联系将有一个深入的认识，将为重症医学的研究在理论上有一个大的跨跃；本课题采用最新的蛋白质组学的先进方法和技术，从组织病理、细胞和分子水平以及临床验证多个层面展开基础和临床研究，具有系统、全面的特色。