TRPV1-dependent neuro-immune modulation and regulation of endogenous acyl-dopamines in sepsis and acute inflammation

项目摘要 脓毒症是一个可怕的公共卫生问题，需要更好地了解其基本机制 脓毒症和免疫调节我们的研究将确定抗炎机制和功能 内源性N-油酰多巴胺（OLDA）和N-花生四烯酸多巴胺（NADA）在脓毒症中的作用 在外周和中枢神经系统（PNS，CNS）中产生的酰基多巴胺。OLDA和NADA展览 瞬时受体电位香草素1（TRPV 1）和大麻素受体1（CB 1 R）的活性。小鼠 S.金黄色葡萄球菌肺炎、腹腔内多微生物败血症或内毒素血症，我们发现OLDA或NADA 给药1）诱导全身性IL-10的早期显著上调，2）降低促炎性因子， 细胞因子和趋化因子，和3）在24小时降低急性肺损伤和脓毒症评分。NADA也 降低CGRP并增加LPS处理小鼠血浆中的P物质。我们对骨髓的研究 Trpv 1-/-和野生型小鼠的嵌合体表明，NADA通过非- 骨髓细胞我们进一步的初步数据表明，神经元TRPV 1介导了抗炎作用， OLDA在LPS处理的小鼠中的保护作用，更具体地说，这些作用是由 TRPV 1神经元在CNS而不是PNS中。最后，我们观察到血浆和大脑中的 OLDA和NADA在LPS处理的小鼠中，这表明这些新的内源性脂质可能是动态的， 在急性炎症和脓毒症期间调节。总的来说，我们的数据导致了我们的中心假设， 存在一种基于CNS的神经免疫调节联系，其涉及神经元TRPV 1的激活， 内源性脂质，如OLDA和NADA，并调节炎症反应和急性 炎症和败血症。目的#1将定位负责抗炎作用的TRPV 1神经元 OLDA和NADA在急性炎症和脓毒症中的作用，并确定CB 1 R在TRPV 1依赖性 OLDA和NADA的抗炎作用。目标#2将确定下游机制， OLDA或NADA激活神经元TRPV 1可上调外周血单核细胞产生IL-10 脓毒症，重点是P物质，CGRP和交感神经系统的作用。目标#3将定义 OLDA和NADA对脓毒症结局的影响，以及脓毒症对内源性 OLDA和NADA以及TRPV 1和CB 1 R的表达。这些研究将促进对 脓毒症中NADA、OLDA和TRPV 1的神经免疫调节作用和内源性作用， 最终目标是确定新的治疗靶点。

PROJECT SUMMARY Sepsis is a formidable public health problem, and there is a need to better understand the basic mechanisms of sepsis and immunomodulation. Our studies will define the anti-inflammatory mechanisms and functional effects in sepsis of N-oleoyl dopamine (OLDA) and N-arachidonoyl dopamine (NADA), which are endogenous acyl-dopamines produced in the peripheral and central nervous systems (PNS, CNS). OLDA and NADA exhibit activity at the transient receptor potential vanilloid 1 (TRPV1) and cannabinoid receptor 1 (CB1R). In mice with S. aureus pneumonia, intraabdominal polymicrobial sepsis, or endotoxemia, we found that OLDA or NADA administration 1) induces an early substantial upregulation of systemic IL-10, 2) decreases pro-inflammatory cytokines and chemokines, and 3) reduces acute lung injury and sepsis scores at 24 hours. NADA also reduces CGRP and increases Substance P in the plasmas of LPS-treated mice. Our studies in bone marrow chimeras of Trpv1-/- and wild-type mice indicated that NADA induces IL-10 via TRPV1 expressed by non- myeloid cells. Our further preliminary data suggest that neuronal TRPV1 mediates the anti-inflammatory and protective actions of OLDA in LPS-treated mice, and more specifically, that these effects are mediated by TRPV1 neurons in the CNS rather than the PNS. Finally, we observed increased plasma and brain levels of OLDA and NADA in LPS-treated mice, which suggest that these novel endogenous lipids may be dynamically regulated during acute inflammation and sepsis. Collectively, our data have led to our central hypothesis that there is a CNS based neuro-immunomodulatory link that involves the activation of neuronal TRPV1 by endogenous lipids, such as OLDA and NADA, and regulates inflammatory responses and outcomes of acute inflammation and sepsis. Aim #1 will localize the TRPV1 neurons responsible for the anti-inflammatory actions of OLDA and NADA in acute inflammation and sepsis and determine the role of CB1R in the TRPV1-dependent anti-inflammatory actions of OLDA and NADA. Aim #2 will identify the downstream mechanisms by which the activation of neuronal TRPV1 by OLDA or NADA upregulates IL-10 production by circulating monocytes in sepsis, focusing on the roles of Substance P, CGRP, and the sympathetic nervous system. Aim #3 will define the effects of OLDA and NADA on sepsis outcomes, and the effects of sepsis on endogenous production of OLDA and NADA, and expression of TRPV1 and CB1R. These studies will advance the understanding of the neuro-immunomodulatory effects and endogenous roles of NADA, OLDA and TRPV1 in sepsis, with the ultimate goal to identify novel therapeutic targets.