摘要目的:探讨脓毒症小鼠不同时间点免疫相关器官组织c-Jun氨基末端激酶（JNK）、CCAAT/增强子结合蛋白同源蛋白（CHOP）凋亡状况及细胞因子水平变化。方法:采用区组随机法将27只雄性BALB/c小鼠分为正常组、脓毒症6 h组、脓毒症12 h组，每组9只。采用盲肠结扎穿孔术（CLP）制备脓毒症模型。各组分别于相应时间点取血，采用酶联免疫吸附试验（ELISA）检测血清肿瘤坏死因子-α（TNF-α）、白细胞介素（IL-1β、IL-10）等细胞因子水平；取血后处死小鼠取脾脏、胸腺、阑尾组织，采用蛋白质免疫印迹试验（Western Blot）检测磷酸化JNK（p-JNK）、JNK1、CHOP、活化的天冬氨酸特异性半胱氨酸蛋白酶3（caspase-3）蛋白表达。结果:与正常组比较，脓毒症6 h组细胞因子水平和脾脏组织p-JNK/JNK1比值、CHOP、caspase-3表达量以及胸腺、阑尾组织CHOP、caspase-3表达量均显著升高〔血TNF-α（ng/L）：24.29±3.09比2.93±2.09，血IL-1β（ng/L）：5.00±3.19比3.54±1.53，血IL-10（ng/L）：1 963.93±270.20比275.09±45.21，脾脏p-JNK/JNK1比值：0.257±0.126比0.154±0.068，脾脏CHOP/β-actin：0.201±0.131比0.142±0.068，脾脏caspase-3/β-actin：0.215±0.126比0.098±0.088，胸腺CHOP/β-actin：0.122±0.071比0.089±0.067，胸腺caspase-3/β-actin：0.258±0.145比0.108±0.045，阑尾CHOP/β-actin：0.361±0.134比0.215±0.112，阑尾caspase-3/β-actin：0.439±0.211比0.321±0.145，均 P<0.05〕，而胸腺、阑尾组织p-JNK/JNK1比值差异无统计学意义（胸腺p-JNK/JNK1比值：1.221±0.776比1.168±0.475，阑尾p-JNK/JNK1比值：2.014±1.227比1.828±0.915，均 P>0.05）。脓毒症12 h组除IL-10水平明显下降外，其他细胞因子水平及脾脏、胸腺、阑尾组织p-JNK/JNK1比值、CHOP、caspase-3表达量均较脓毒症6 h组进一步升高〔血IL-10（ng/L）：1 698.98±210.52比1 963.93±270.20，血TNF-α（ng/L）：41.66±6.57比24.29±3.09，血IL-1β（ng/L）：10.37±4.14比5.00±3.19，脾脏p-JNK/JNK1比值：0.399±0.135比0.257±0.126，脾脏CHOP/β-actin：0.298±0.145比0.201±0.131，脾脏caspase-3/β-actin：0.353±0.145比0.215±0.126，胸腺p-JNK/JNK1比值：1.667±0.891比1.221±0.776，胸腺CHOP/β-actin：0.207±0.133比0.122±0.071，胸腺caspase-3/β-actin：0.416±0.179比0.258±0.145，阑尾p-JNK/JNK1比值：2.425±1.361比2.014±1.227，阑尾CHOP/β-actin：0.456±0.189比0.361±0.134，阑尾caspase-3/β-actin：0.635±0.289比0.439±0.211，均 P<0.05〕。 结论:内质网途径JNK和CHOP通路参与小鼠脓毒症时免疫相关细胞凋亡及细胞因子表达，脓毒症12 h较6 h时凋亡更加明显，炎症反应更强。

abstractsObjective:To investigate the c-Jun N-terminal kinase (JNK), CCAAT/enhancer-binding protein homologous protein (CHOP) pathway apoptosis and the changes of cytokine levels in immune-related organs and tissues of sepsis mice at different time points.Methods:Twenty-seven male BALB/c mice were divided into normal group, sepsis 6 hours group and sepsis 12 hours group by the block randomization method, with 9 mice in each group. The sepsis model was made by cecal ligation and puncture (CLP). Blood sample was collected from each group at the corresponding time point, and the serum levels of tumor necrosis factor-α (TNF-α), interleukin (IL-1β, IL-10) were measured by enzyme linked immunosorbent assay (ELISA). The spleen, thymus and appendix tissues were taken from the mice to detect the expressions of phosphorylation-JNK (p-JNK), JNK1, CHOP and cleaved caspase-3 protein by Western Blot.Results:The level of cytokines, p-JNK/JNK1 ratio, CHOP and caspase-3 in spleen tissues, and the CHOP, caspase-3 in thymus and appendix tissue in the sepsis 6 hours group were significantly higher than those in the normal group [serum TNF-α (ng/L): 24.29±3.09 vs. 2.93±2.09, serum IL-1β (ng/L): 5.00±3.19 vs. 3.54±1.53, serum IL-10 (ng/L): 1 963.93±270.20 vs. 275.09±45.21, spleen p-JNK/JNK1 ratio: 0.257±0.126 vs. 0.154±0.068, spleen CHOP/β-actin: 0.201±0.131 vs. 0.142±0.068, spleen caspase-3/β-actin: 0.215±0.126 vs. 0.098±0.088, thymus CHOP/β-actin: 0.122±0.071 vs. 0.089±0.067, thymus caspase-3/β-actin: 0.258±0.145 vs. 0.108±0.045, appendix CHOP/β-actin: 0.361±0.134 vs. 0.215±0.112, appendix caspase-3/β-actin: 0.439±0.211 vs. 0.321±0.145, all P < 0.05]. However, there were no significant difference in the p-JNK/JNK1 ratio in thymus and appendix (thymus p-JNK/JNK1 ratio: 1.221±0.776 vs. 1.168±0.475, appendix p-JNK/JNK1 ratio: 2.014±1.227 vs. 1.828±0.915, both P > 0.05). Cytokine levels and the p-JNK/JNK1 ratio, CHOP, caspase-3 in spleen, thymus, and appendix in the sepsis 12 hours group were further increased when compared with those in the sepsis 6 hours group, except for a significant decrease in IL-10 level [serum IL-10 (ng/L): 1 698.98±210.52 vs. 1 963.93±270.20, serum TNF-α (ng/L): 41.66±6.57 vs. 24.29±3.09, serum IL-1β (ng/L): 10.37±4.14 vs. 5.00±3.19, spleen p-JNK/JNK1 ratio: 0.399±0.135 vs. 0.257±0.126, spleen CHOP/β-actin: 0.298±0.145 vs. 0.201±0.131, spleen caspase-3/β-actin: 0.353±0.145 vs. 0.215±0.126, thymus p-JNK/JNK1 ratio: 1.667±0.891 vs. 1.221±0.776, thymus CHOP/β-actin: 0.207±0.133 vs. 0.122±0.071, thymus caspase-3/β-actin: 0.416±0.179 vs. 0.258±0.145, appendix p-JNK/JNK1 ratio: 2.425±1.361 vs. 2.014±1.227, appendix CHOP/β-actin: 0.456±0.189 vs. 0.361±0.134, appendix caspase-3/β-actin: 0.635±0.289 vs. 0.439±0.211, all P < 0.05]. Conclusions:The endoplasmic reticulum pathway JNK and CHOP pathways are involved in immune-related cell apoptosis and cytokine expression in mice with sepsis. Apoptosis is more obvious at 12 hours than at 6 hours, and the inflammatory response is stronger.