Mechanisms of Novel Herbal Therapies for Sepsis.

描述（由申请人提供）：尽管最近在抗生素治疗和重症监护方面取得了进展，但脓毒症仍然是重症监护病房中最常见的死亡原因，仅在美国每年就有> 225，000名受害者。其发病机制部分归因于由早期促炎细胞因子（例如，TNF和IFN-γ），但通过迟效促炎介质（例如，HMGB1）。靶向早期促炎细胞因子的药剂（例如，TNF）可以是保护性的，如果给药，而能够抑制HMGB 1释放或活动的药物可以拯救动物免于致命的脓毒症，即使在疾病发作后给予。我们开创性地发现HMGB 1作为致死性脓毒症的晚期介质，这促使了对开发新的实验治疗方法的进一步研究。我们已经产生了初步的数据表明，几种常用的中药，丹参（丹参，类固醇样丹参酮）和绿色茶（茶树，表没食子儿茶素没食子酸酯，EGCG）的主要成分有效地衰减内毒素诱导的HMGB 1释放，并提高动物的存活率在小鼠模型的内毒素血症和败血症时，腹腔内给药。然而，尚不清楚草药成分单独或组合是否以及如何影响由其他炎症刺激（例如，G+细菌外毒素、CpG-DNA、TNF或IFN-γ），并因此影响致死性内毒素血症和脓毒症的结果，如果通过临床上可行的（静脉内或口服）给药途径给药。目的1中概述的实验将检验草药成分影响由其他外源性（例如，G+外毒素或CpG-DNA）或内源性（例如，TNF或IFN-γ）刺激，并且草药成分分散地影响HMGB 1诱导的一氧化氮、趋化因子和生长因子的释放。在目标2中，我们将测试一种新的假设，即草药成分通过促进细胞外HMGB 1的内吞“再摄取”（再循环）或通过刺激自噬HMGB 1降解来抑制HMGB 1释放。目的3中概述的实验将检验以下假设：口服或静脉内施用草药组分通过调节腹膜白细胞浸润、全身性和腹膜炎症、组织损伤和器官功能障碍来保护动物免于致死性内毒素血症和脓毒症。这些问题的答案将显着提高我们对两种常用中药的免疫调节机制的理解，并为脓毒症和其他炎症性疾病的治疗提供替代策略。 公共卫生相关性：脓毒症仍然是重症监护病房中最常见的死亡原因，仅在美国每年就有超过225，000名受害者。它是一种多因素疾病，引发不受控制的全身炎症反应，最终导致多器官衰竭和死亡。我们最近发现HMGB 1作为实验性脓毒症的晚期介质，促使中草药作为潜在的HMGB 1靶向治疗剂的研究。本申请的长期目标是阐明两种草药[丹参（Salvia miltiorrhiza））和绿色茶（Camellia sinensis）]的主要成分抑制HMGB 1释放的新机制，从而保护动物免受致死性内毒素血症和脓毒症。

DESCRIPTION (provided by applicant): Despite recent advance in antibiotic therapy and intensive care, sepsis remains the most common cause of death in the intensive care units, claiming > 225,000 victims annually in the U.S. alone. Its pathogenesis is partly attributable to dys-regulated inflammatory responses that are propagated by early proinflammatory cytokines (e.g., TNF and IFN-gamma) but sustained by late-acting proinflammatory mediators (e.g., HMGB1). Agents targeting early proinflammatory cytokines (e.g., TNF) could be protective if given prophylatically; whereas agents capable of inhibiting HMGB1 release or activities could rescue animals from lethal sepsis even if given after onset of disease. Our seminal discovery of HMGB1 as a late mediator of lethal sepsis has prompted further investigation for developing new experimental therapeutics. We have generated preliminary data indicating that major components of several commonly used Chinese herbs, Danshen (Salvia miltiorrhiza, steroid-like tanshinones) and Green tea (Camellia sinensis, epigallocatechin gallate, EGCG) effectively attenuated endotoxin-induced HMGB1 release, and improved animal survival in murine models of endotoxemia and sepsis when given intraperitoneally. However, it is not known whether and how herbal components, individually or in combination, affect HMGB1 release induced by other inflammatory stimuli (e.g., G+ bacterial exotoxin, CpG-DNA, TNF, or IFN-gamma), and consequently influence the outcome of lethal endotoxemia and sepsis if given via clinically feasible (intravenous or oral) route of administration. The experiments outlined in Aim 1 will test the hypothesis that herbal components affect HMGB1 release induced by other exogenous (e.g., G+ exotoxin or CpG-DNA) or endogenous (e.g., TNF or IFN-gamma) stimuli, and that herbal components divergently influence HMGB1-induced release of nitric oxide, chemokines, and growth factors. In Aim 2, we will test a novel hypothesis that herbal components inhibit HMGB1 release either by facilitating endocytic "re-uptake" (recycling) of extracellular HMGB1, or by stimulating autophagic HMGB1 degradation. The experiments outlined in Aim 3 will test the hypothesis that oral or intravenous administration of herbal components protects animals against lethal endotoxemia and sepsis by modulating peritoneal leukocyte infiltration, systemic and peritoneal inflammation, tissue injury, and organ dysfunction. Answers to these questions will significantly improve our understanding of immune modulatory mechanisms of two commonly used Chinese medicinal herbs, and shed light on the development of alternative strategies for treatment of sepsis and other inflammatory diseases. PUBLIC HEALTH RELEVANCE: Sepsis remains the most common cause of death in the intensive care units, claiming > 225,000 victims annually in the U.S. alone. It is a multi-factorial disorder that triggers an uncontrolled systemic inflammatory response, ultimately leading to multiple organ failure and death. Our recent discovery of HMGB1 as a late mediator of experimental sepsis has prompted investigation of Chinese medicinal herbs as potential HMGB1- targeting therapeutic agents. The long-term goal of this application is to elucidate novel mechanisms by which major components of two medicinal herbs [Danshen (Salvia miltiorrhiza) and Green tea (Camellia sinensis)] inhibit HMGB1 release, and consequently protect animals against lethal endotoxemia and sepsis.