Mechanisms of Novel Herbal Therapies for Sepsis.

脓毒症新型草药疗法的机制。

基本信息

批准号：
10663911
负责人：
Haichao Wang
金额：
$ 41.88万
依托单位：
FEINSTEIN INSTITUTE FOR MEDICAL RESEARCH
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-09-01 至 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10663911
关键词：
Affect Age Animal Model Animals Anti-Inflammatory Agents Antibiotic Therapy Antibodies Bacteremia Bacterial Infections Biological Markers Blood Blood specimen Cause of Death Circulation Clinical Coagulation Process Collection Connexin 43 Data Diagnosis Distant Dose Double-Stranded RNA Endotoxemia Fasciola hepatica Gender Genes Genetic HMGB1 gene Herbal Medicine Human Immune Immunoglobulin G Impairment In Vitro Inflammation Inflammatory Inflammatory Response Intensive Care Intensive Care Units Interleukin-1 beta Interleukin-10 Knock-out Lanosterol Lead Light Lung infections Lycium Macrophage Mediating Mediator Medicinal Plants Mus Natural Products Outcome PRKR gene Parasites Pathogenesis Pathogenicity Pathologic Patients Pharmaceutical Preparations Phosphorylation Phytotherapy Progesterone Proteins Recombinants Role Sepsis Supplementation Surrogate Markers Survival Rate TLR4 gene TNF gene Testing Time cecal ligation puncture chemokine cytokine experimental study high throughput screening human subject improved in vivo inhibitor monocyte neutralizing antibody novel protective effect protein kinase R secondary metabolite septic septic patients systemic inflammatory response

项目摘要

Project Abstract Despite recent advances in antibiotic therapy and intensive care, sepsis remains the most common cause of death in the intensive care unit, annually claiming >225,000 victims in the U.S. alone. The pathogenesis of sepsis remains obscure, but is partly attributable to dys-regulated inflammation propagated by “early” cytokines (e.g., TNF and IL-1β), but sustained by “intermediate” (e.g., SAA) and “late” (e.g., HMGB1) mediators. We recently discovered that LPS and SAA upregulated the expression of connexin 43 (Cx43) and/or pannexin 1 (Panx1) hemichannels to facilitate ATP-dependent PKR activation and HMGB1 release, but did not know whether LPS and SAA also induced procathepsin L (pCTS-L) secretion to mediate lethal bacterial infections (LBI). Our preliminary data indicated that LPS and SAA induced a marked expression and secretion of pCTS- L in both murine macrophage and human monocyte cultures. Consequently, pCTS-L was not detectable in the circulation of healthy animals or human subjects, but significantly elevated in the blood of septic animals and patients. Highly purified recombinant pCTS-L stimulated primary human monocytes to release various chemokines, as well as pro- (e.g., TNF and IL-1β) and anti-inflammatory cytokines (e.g., IL-10) in vitro, and exacerbated endotoxemic lethality in vivo. In contrast, pCTS-L-neutralizing antibodies significantly rescued mice from lethal sepsis, suggesting pCTS-L as another late mediator of LBI. Meanwhile, a semi-high throughput screening of a NatProduct Collection of 800 natural products and a US Drug Collection of 1360 bioactive compounds led to the finding of a few lead compounds [including lanosterol (LAN) and progesterone (PRO)] with striking structural resemblance and similar pCTS-L-inhibiting activities. Although LAN is an abundant secondary metabolite in some medicinal plants including wolfberry and aveloz, it also serves as a substrate for the synthesis of PRO in animals and humans. These exciting findings prompted the current proposal to investigate a novel role of pCTS-L in LBI, as well as intricate mechanisms by which a natural product, LAN, and its derivative, PRO, inhibit pCTS-L-induced inflammation. The experiments outlined in Aim 1 will test the hypothesis that pCTS-L systemically accumulates in the circulation of septic patients and correlates with other surrogate markers of sepsis. In Aim 2, we will test the hypothesis that alterations of pCTS-L levels (by supplementation of pCTS-L or genetic knockout) or activities (by use of neutralizing IgGs or natural inhibitor LAN or PRO) divergently influence the outcomes of LBI. The experiments outlined in Aim 3 will test the hypothesis that LAN and/or PRO inhibit the pCTS-L-induced inflammation through impairing the TLR4/ RAGE-dependent hemichannel and PKR activation in vitro, and confer protection against LBI partly by attenuating systemic inflammation and associated dysregulated coagulation in vivo. Collectively, this project will improve our understanding of the role of pCTS-L in LBI, and shed light on the intricate mechanism underlying natural product LAN and PRO-mediated protection against lethal bacterial infections.

项目摘要尽管抗生素治疗和重症监护最近取得了进展，但败血症仍然是最常见的原因仅在美国，每年就有超过 225,000 名患者在重症监护室死亡。脓毒症仍然不清楚，但部分归因于“早期”传播的失调炎症细胞因子（例如 TNF 和 IL-1β），但由“中间”（例如 SAA）和“晚期”（例如 HMGB1）介质维持。我们最近发现 LPS 和 SAA 上调连接蛋白 43 (Cx43) 和/或 pannexin 的表达 1 (Panx1) 半通道促进 ATP 依赖性 PKR 激活和 HMGB1 释放，但不知道 LPS 和 SAA 是否也诱导组织蛋白酶 L (pCTS-L) 分泌以介导致命的细菌感染 (LBI)。我们的初步数据表明LPS和SAA诱导pCTS-的显着表达和分泌。 L 在小鼠巨噬细胞和人单核细胞培养物中均未检测到。健康动物或人类受试者的血液循环，但在脓毒症动物的血液中显着升高，高度纯化的重组 pCTS-L 刺激原代人单核细胞释放各种物质。趋化因子，以及体外促细胞因子（例如 TNF 和 IL-1β）和抗炎细胞因子（例如 IL-10），以及相比之下，pCTS-L-中和抗体显着挽救了体内内毒素致死性。小鼠致死性脓毒症，表明 pCTS-L 是 LBI 的另一种晚期介质。对 NatProduct Collection 的 800 种天然产物和 US Drug Collection 的 1360 种天然产物进行通量筛选生物活性先导化合物发现了一些先导化合物[包括羊毛甾醇 (LAN) 和黄体酮 (PRO)] 具有惊人的结构相似性和类似的 pCTS-L 抑制活性。在一些药用植物（包括枸杞和阿维洛兹）中含有丰富的次生代谢产物，它也可以作为在动物和人类中合成 PRO 的底物这些令人兴奋的发现促使了当前的研究。提议研究 pCTS-L 在 LBI 中的新作用，以及天然药物的复杂机制产品 LAN 及其衍生物 PRO 可抑制 pCTS-L 诱导的炎症。目标 1 将检验 pCTS-L 在脓毒症患者循环系统中蓄积的假设，以及与脓毒症的其他替代标志物相关。在目标 2 中，我们将检验以下假设： pCTS-L 水平（通过补充 pCTS-L 或基因敲除）或活性（通过使用中和 IgG 或天然抑制剂 LAN 或 PRO）对 LBI 的结果产生不同的影响目标 3 中概述的实验。将检验以下假设：LAN 和/或 PRO 通过损害 TLR4/RAGE 依赖性半通道和 PKR 体外激活，并部分通过以下方式提供针对 LBI 的保护：总的来说，该项目可以减轻全身炎症和相关的体内凝血失调。将提高我们对 pCTS-L 在 LBI 中的作用的理解，并阐明复杂的机制天然产物 LAN 和 PRO 介导的针对致命细菌感染的保护作用。