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目的:通过观察脑细胞程序性坏死和脑组织中90个细胞因子水平的变化，探讨心搏骤停心肺复苏（CPR）后大鼠脑损伤的分子机制。方法:按随机数字表法将SD大鼠分为假手术组（Sham组，n n＝10）和心搏骤停组（n n＝10）。采用窒息法诱导大鼠心搏骤停，并于心搏骤停6 min后开始CPR；Sham组仅常规行气管插管等，未诱导心搏骤停。CPR后3 d对大鼠进行神经功能缺损评分（NDS）后取血并处死大鼠，采用酶联免疫吸附试验（ELISA）检测血清S100B水平，用免疫荧光法检测脑组织程序性坏死细胞，用蛋白芯片检测脑组织90个细胞因子表达水平，以两组蛋白的相对比值≥1.5或≤0.5且n P<0.05为差异表达蛋白。n 结果:心搏骤停组有8只大鼠成功复苏，死亡2只，为使样本量匹配，Sham组随机选择8只进行实验。与Sham组比较，心搏骤停组大鼠NDS评分明显降低〔分：63.0（62.5，64.3）比80.0（80.0，80.0），n P<0.01〕，血清S100B水平明显升高（ng/L：47.96±10.16比16.56±5.60，n P<0.01）。心搏骤停后脑皮质和海马区程序性坏死细胞较Sham组多见n 〔原位末端缺刻标记法（TUNEL）阳性且天冬氨酸特异性半胱氨酸蛋白酶3（caspase-3）阴性的细胞比例：脑皮质为（15.70±0.32）%比（8.00±0.28）%，海马区为（20.80±1.35）%比（9.00±4.00）%，均n P<0.05〕。蛋白芯片检测显示，与Sham组比较，心搏骤停组脑组织炎性相关细胞因子细胞因子诱导中性粒细胞趋化因子（CINC-2α/β、CINC-3）、γ-干扰素（IFN-γ）和信号蛋白C -端Src激酶（CSK）表达水平明显升高（心搏骤停组与Sham组蛋白表达比值：CINC-2 α/β为2.503±0.428，n P＝0.024；CINC-3为2.369±0.142，n P＝0.005；IFN-γ为3.149±1.362，n P＝0.044；CSK为1.887±0.105，n P＝0.001），神经保护因子睫状神经营养因子（CNTF）、胶质细胞源性神经营养因子受体（GFRα-1、GFRα-2）、生长激素（GH）、生长激素受体（GHR）、粒-巨噬细胞集落刺激因子（GM-CSF）以及抗炎细胞因子白细胞介素- 10（IL-10）表达水平明显降低n （心搏骤停组与Sham组蛋白表达比值：CNTF为0.341±0.137，n P＝0.036；GFRα-1为0.461±0.164，n P＝0.044；GFRα-2为0.447±0.017，n P＝0.033；GH为0.450±0.136，n P＝0.024；GHR为0.508±0.128，n P＝0.022；GM-CSF为0.446±0.130，n P＝0.035；IL-10为0.502±0.211，n P＝0.017）。n 结论:程序性坏死参与心搏骤停CPR后大鼠脑损伤，脑损伤分子机制可能与炎症反应、神经再生障碍和神经细胞存活受损有关。“,”Objective:To investigate the underlying molecular mechanisms of brain injury in rats after cardiac arrest and cardiopulmonary resuscitation (CPR) by observing necroptosis of brain cells and changes of 90 cytokines in brain tissue.Methods:Sprague-Dawley (SD) rats were divided into Sham group (n n = 10) and cardiac arrest group (n n = 10) according to random number table method. The model of asphyxia cardiac arrest for 6 minutes followed by CPR model was established. Tracheal intubation in Sham rats were routinely performed without inducing cardiac arrest. Neurological deficit score (NDS) was evaluated, blood samples were collected and rats were sacrificed, then serum S100B level was measured by enzyme linked immunosorbent assay (ELISA) on the third day after CPR. Necroptotic cells in brain were detected by immunofluorescence staining. The levels of 90 cytokines expression in brain were measured by antibody array. The relative ratio of the two groups of protein expression ≥ 1.5 or ≤ 0.5 and n P < 0.05 represented the differential expression protein.n Results:There were 8 rats successfully resuscitated and 2 died in cardiac arrest group. There were 8 rats selected in Sham group to match the sample size. Compared with Sham group, the NDS score of cardiac arrest group was significantly lower [63.0 (62.5, 64.3) vs. 80.0 (80.0, 80.0), n P < 0.01], and the level of serum S100B was significantly higher (ng/L: 47.96±10.16 vs. 16.56±5.60, n P < 0.01). More necroptotic cells in cerebral cortex and hippocampus were found in cardiac arrest group [proportion of cells positive for TdT-mediated nick end labeling (TUNEL) and negative for caspase-3: (15.70±0.32)% vs. (8.00±0.28)% in cortex, (20.80±1.35)% vs. (9.00±4.00)% in hippocampus, both n P < 0.05]. The levels of inflammatory cytokines [cytokine-induced neutrophil chemoattractant (CINC-2α/β, CINC-3), interferon-γ (IFN-γ)] and signal protein c-Src kinase (CSK) in brain significantly increased after cardiac arrest as compared to Sham group levels (ratio of cardiac arrest group to Sham group: CINC-2 α/β was 2.503±0.428, n P = 0.024; CINC-3 was 2.369±0.142, n P = 0.005; IFN-γwas 3.149±1.362,n P = 0.044; CSK was 1.887±0.105, n P = 0.001). However, the levels of neuroprotective cytokines ciliary neurotrophic factor (CNTF), glial cell-derived neurotrophic factor receptor (GFR α-1, GFR α-2), growth hormone (GH), growth hormone receptor (GHR), granulocyte-macrophage colony-stimulating factor (GM-CSF) and anti-inflammatory protein interleukin-10 (IL-10) significantly decreased after cardiac arrest (ratio of cardiac arrest group to Sham group: CNTF was 0.341±0.137, n P = 0.036; GFRα-1 was 0.461±0.164,n P = 0.044; GFRα-2 was 0.447±0.017,n P = 0.033; GH was 0.450±0.136, n P = 0.024; GHR was 0.508±0.128, n P = 0.022; GM-CSF was 0.446±0.130, n P = 0.035; IL-10 was 0.502±0.211, n P = 0.017).n Conclusions:Necroptosis is involved in brain injury after cardiac arrest. The molecular mechanisms of brain injury may be related to inflammatory response, neurogenesis disorder and impaired survival of nerve cells.