基于宽场显微荧光成像技术的一氧化氮亚硝基化修饰对激活的嗜中性粒细胞质钙调节机制的研究

项目名称： 基于宽场显微荧光成像技术的一氧化氮亚硝基化修饰对激活的嗜中性粒细胞质钙调节机制的研究

项目编号： No.11204142

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 物理学I

项目作者： 潘雷霆

作者单位： 南开大学

项目金额： 28万元

中文摘要： 嗜中性粒细胞是机体抵御外界病原侵入的第一道防线，在各种免疫反应过程中发挥着重要作用。研究表明其一般具有静息和激活两种状态。我们已报道一氧化氮（Nitric Oxide,NO）作为生物体内著名的"明星分子"可通过巯基亚硝基化修饰方式诱导激活的人嗜中性粒细胞钙库控制的钙内流（Store-operated Calcium Entry,SOCE）。本项目拟采用宽场显微荧光成像技术，研究NO亚硝基化修饰对激活的人嗜中性粒细胞钙的详细调节机制：阐明NO亚硝基修饰诱导激活的嗜中性粒细胞SOCE 过程的信号转导通路；实现胞质钙荧光双通道快速比率成像和胞质钙、氢离子双通道同步监测；有效获取NO亚硝基化修饰诱导的SOCE 过程快速时空偶联信息；揭示激活与静息态的嗜中性粒细胞质钙对NO亚硝基修饰不同响应的关键机制。本研究可为临床NO治疗嗜中性粒细胞激活导致的冠脉循环堵塞等疾病提供有效的实验支持和新的思考视角。

中文关键词： 嗜中性粒细胞；亚硝基化修饰；一氧化氮；钙；荧光成像

英文摘要： As the first line of host defense against invasion of pathogenic microbes, neutrophils play a crucial role in a variety of inflammatory responses. In generally, researchers have known that there are two main states for neutrophils: resting and activation. Our previous work reported that nitric oxide (NO), a new super "star molecule" of organism, could induce store-operated calcium entry through (SOCE) via s-nitrosylation modification in activation-dependent human neutrophils. In this proposal, we intend to investigate the detailed mechanism of NO on cytosolic calcium via s-nitrosylation modification in activated neutrophils based on wide-field fluorescence microscopy. The main content of this studies occurs as follows: clarifying the signal transduction pathways of s-nitrosylation modification-induced SOCE in activated human neutrophils; achieving simultaneous imaging of calcium and hydrogen & high speed two-channel ratio imaging of calcium in activated neurtrophils; acquiring fast spatiotemporal information of s-nitrosylation modification-induced SOCE in activated neutrophils; understanding the crucial points of s-nitrosylation modification-induced SOCE in activated neutrophils which is different to that of resting neutrophils. This study on modulation of neutrophils calcium by exogenous NO might be much more u

英文关键词： Neutrophils；s-nitrosylation Modification；Nitric Oxide；Calcium；Fluorescence Imaging