CO中毒是当今发生率和死亡率最高的中毒，急性CO中毒迟发性脑病（DEACMP）是CO中毒后发病率、致残率和致死率较高的并发症，但其发病机制尚未完全阐明。研究发现免疫炎症机制在其发生发展中发挥着更重要的作用，与我们前期研究CO中毒后炎症因子攻击使脑内髓鞘碱性蛋白脱失一致。小胶质细胞是中枢神经系统主要的免疫细胞，CO中毒后可被激活，其机制尚无报道。文献报道胞外ATP介导小胶质细胞参与免疫应答，小胶质细胞上P2Y12受体激活与炎症因子分泌密切相关。本研究拟建立DEACMP模型并给予P2Y12受体拮抗剂，应用水迷宫实验评价认知功能，利用高效液相色谱法、流式细胞仪、荧光双标检测海马组织内ATP含量、活化小胶质细胞及P2Y12受体表达，阐明DEACMP的发病机制，为该病防治提供新的思路和依据。

Delayed encephalopathy after acute carbon monoxide poisoning (DEACMP) is one of the complications after carbon monoxide (CO) poisoning, with high incidence, morbidity and mortality, while the underlying mechanism remains elusive. Studies have demonstrated that inflammatory immune mechanism might play a crucial role in the pathogenesis, as same as that demyelination with myelin basic protein(MBP) decreased after CO poisoning due to the attack of inflammatory factors in our previous study. Microglia as been widely known to be the main immune cell of central nervous system, can be activated by CO poisoning, however, the activation mechanism of microglia has not been reported yet. Microglia cells activated by extracellular ATP may participate in the immune response to brain injury, and the activation of P2Y12 on microglia was associated with the expression of some inflammatory factors. In this study, firstly, DEACMP model was established after CO poisoning. Then injected the P2Y12 receptor agonist , at different time point, water maze was used to evaluate cognitive function, high performance liquid chromatography to determine the ATP content in hippocampucs, flow cytometer to examine the activated microglia cells, double-label immunofluorescence stain to observe the localization of microglia and P2Y12 receptor. This study not only would investigate the pathogenesis mechanism of DEACMP, but also provide new information to understand of the treatment of DEACMP in future clinic practice.

一氧化碳（CO）中毒是常见的中毒之一，中毒后可造成以中枢神经系统功能损害为主的多脏器病变，其中急性一氧化碳中毒迟发型脑病（ DEACMP）是最常见且严重的并发症，发病率约为0.2%～47%，死亡率可达31%。目前DEACMP发病机制尚未完全阐明，且无有效的治疗手段及策略。为进一步探讨其发病机制，本研究建立了符合临床特点的DEACMP动物模型；应用高效液相色谱法检测海马组织内ATP 的含量；应用流式细胞仪检测海马组织内小胶质细胞活化情况；应用免疫荧光双标技术检测小胶质细胞及P2Y12 受体的表达情况。研究结果显示：中毒后大鼠出现典型CO中毒表现，碳氧血红蛋白（C0Hb）含量与吸入CO浓度呈显著正相关(r=0.748，P<0．05),1000ppm、3000ppm、4000ppm组各组死亡率分别为0% 、48.39 % 、58.97 % ；结合Morris水迷宫检测大鼠认知功能下降、病理结果观察海马组织有细胞损伤判断DEACMP发生情况: 各实验组DEACMP发生率分别为6.25% 、28.13% 、31.25%，其中在中毒后21d、28d发生DEACMP的大鼠数量明显增加，21d、28d组间比较差异无统计学意义，选择死亡率低、发病率高即中毒浓度3000ppm，时间21d为建立DEACMP大鼠模型的最佳方法；在DEACMP高发时间点21d时，大鼠海马组织ATP含量、小胶质细胞活化情况比较结果均为未干预组＞P2Y12受体拮抗剂组＞对照组，差异均有统计学意义（P＜0.05）；免疫荧光双标染色发现静息态小胶质细胞为小胞体，长分枝形态；激活后的小胶质细胞胞体增大，突起回缩，与巨噬细胞形态相似，P2Y12受体在小胶质细胞上可见部分表达，但与预期研究结果存在差异。本研究成功建立了 DEACMP 动物模型，为后续进一步研究DEACMP发病机制及治疗提供了可靠基础；研究发现了DEACMP大鼠小胶质细胞活化水平升高，并发现P2Y12 受体在介导小胶质细胞活化中起到一定作用，P2Y12受体拮抗剂对该病的发生有一定的预防作用，以上研究成果为该病的发病机制、预防及治疗提供了新的思路，对临床早期干预和降低迟发型脑病事件的发生具有重要的临床意义。