中性粒细胞TRPM2通道在脓毒症细菌清除中的作用及机制

项目名称： 中性粒细胞TRPM2通道在脓毒症细菌清除中的作用及机制

项目编号： No.81501702

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 医药、卫生

项目作者： 钱小伟

作者单位： 温州医科大学

项目金额： 18万元

中文摘要： 免疫细胞功能抑制导致脓毒症患者体内细菌清除能力严重下降在脓毒症的发病机制中发挥重要作用。 瞬时受体电位通道M2 (TRPM2) 是高表达于巨噬细胞和PMN的非特异性Ca2+通道。我们前期研究发现TRPM2通道通过促进巨噬细胞杀菌功能保护脓毒症。然而，中性粒细胞 (PMN)TRPM2通道在脓毒症细菌清除中的作用及机制尚不清楚。本研究使用TRPM2-KO小鼠PMN，通过离体杀菌实验和流式细胞检测TRPM2对PMN脱颗粒和杀菌能力的影响；通过杀菌实验、电生理、钙离子测定、流式细胞检测和Western blot等方法深入研究TRPM2对PMN脱颗粒和杀菌作用影响的具体分子机制；使用TRPM2-KO小鼠，通过小鼠脓毒症模型，验证TRPM2通过调控PMN脱颗粒作用促进脓毒症小鼠体内细菌清除保护脓毒症的作用及其机制。本研究将进一步揭示TRPM2保护脓毒症的新机制，为脓毒症的治疗提供新的靶点奠定理论基础。

中文关键词： 瞬时受体电位通道M2；脓毒症；中性粒细胞；细菌清除；系统性全身炎症反应

英文摘要： Increasing evidence supports that immunosuppression exits in septic patients and plays a central role in the pathogenesis of sepsis. The immune cells of both innate and adaptive system are severely suppressed in septic patients, resulting in reduced capability of immune cells to eradicate invading pathogens. Transient Receptor Potential Melastatin 2 (TRPM2), a nonselective Ca2+-permeable channel, is expressed abundantly in macrophages and neutrophils. Our recent study demonstrated a protective role of TRPM2 in controlling bacterial clearance during sepsis possibly by promoting bacterial killing capability of macrophages. However, the role of neutrophil TRPM2 in bacterial clearance during sepsis and its underlying mechanism remain unclear. Using neutrophils from Trpm2-knockout mice, we will firstly investigate the effect of TRPM2 on the bacterial killing capability and degranulation of neutrophils by bacterial killing experiment and flow cytometry. Then, we will further to clarify the detailed molecular mechanism of the effect of TRPM2 on the bacterial killing capability and degranulation of neutrophils by bacterial killing experiment, electrophysiology, intracellular calcium concentration detection, MAPKs inhibitors, flow cytometry, and immunoblot. We will finally confirm the protective role of TRPM2 in promoting bacterial clearance during murine sepsis by regulating the degranulation of neutrophils and its underlying mechanism. Our study will uncover a novel mechanism of the protective role of TRPM2 in sepsis, which provides a new target for the prevention and treatment of sepsis.

英文关键词： Transient Receptor Potential Melastatin 2 ;Sepsis;Neutrophil;Bacterial clearance ;Systemic inflammatory response syndrome