Qiu C F, Wu J F, Pei F, Wang L H, Mei M H, Ouyang B, Guan X D
Department of Critical Care Medicine, First Affiliated Hospital of Sun Yat-sen University, Guangzhou 510080, China.
Zhonghua Yi Xue Za Zhi. 2020 Apr 7;100(13):1033-1037. doi: 10.3760/cma.j.cn112137-20190825-01888.
To observe the effects of 2-aminopurine (2-AP), a double-stranded RNA-dependent protein kinase (PKR) inhibitor, on organ function, plasma inflammatory factor expression and 7 days mortality in sepsis mice induced by cecal ligation puncture (CLP). Forty specific specific pathogen free C57BL/6 mice were randomly divided into sham group (10), CLP group (10), CLP+2-AP group (10) and 2-AP group (10). CLP was used to establish sepsis mice models.Peripheral blood serum was collected 24 hours after operation, alanine aminotransferase (ALT), aspartate aminotransferase (AST), serum creatinine (Cr), blood urea nitrogen (BUN) and inflammatory factors (IL-1β, IL-10 and TNF-α) were detected; peripheral blood and peritoneal lavage fluid were taken for bacterial clearance detection. Another 60 C57BL/6 mice were selected to observe the 7-day survival rate according to the above groups (15). Independent sample test was used to compare the measurement data between groups. The levels of ALT, AST, Cr and BUN in CLP Group and CLP+2-AP group were significantly higher than those in sham group (all 0.001). The levels of ALT and AST in CLP+2-AP group were significantly lower than those in CLP Group (27.88, 11.33, both 0.001); the levels of Cr and BUN in CLP+2-AP group were significantly lower than those in CLP Group (11.02, 7.15, both0.001). Compared with sham group, the levels of pro-inflammatory (IL-1β and TNF-α) and anti-inflammatory (IL-10) cytokines in CLP group were significantly higher (all 0.001); the levels of IL-1β and IL-10 in CLP+2-AP group were significantly lower (all 0.001), but the levels of TNF-α in CLP+2-AP group were not significantly lower (0.33). The 7-day survival rate was 100% in sham group, 13.3% in CLP+2-AP group, 86.7% in 2-AP group and 20.0% in CLP+2-AP group. Inhibition of PKR activation slightly improved the trend of 7-days survival rate of CLP model mice (analysis by mantel Cox test, χ(2)=0.0012, 0.97). In sepsis mice model, inhibition of PKR activity can reduce the expression of inflammatory factors in plasma, decrease bacterial load in blood and abdominal cavity, and protect organ function, which could suggest that inhibition of PKR activity has potential application in sepsis treatment.
观察双链RNA依赖性蛋白激酶（PKR）抑制剂2-氨基嘌呤（2-AP）对盲肠结扎穿刺（CLP）诱导的脓毒症小鼠器官功能、血浆炎症因子表达及7天死亡率的影响。将40只特定病原体-free C57BL/6小鼠随机分为假手术组（10只）、CLP组（10只）、CLP + 2-AP组（10只）和2-AP组（10只）。采用CLP建立脓毒症小鼠模型。术后24小时采集外周血血清，检测丙氨酸氨基转移酶（ALT）、天冬氨酸氨基转移酶（AST）、血清肌酐（Cr）、血尿素氮（BUN）及炎症因子（IL-1β、IL-10和TNF-α）；采集外周血和腹腔灌洗液进行细菌清除检测。另选60只C57BL/6小鼠按上述分组观察7天生存率（每组15只）。采用独立样本检验比较组间计量资料。CLP组和CLP + 2-AP组的ALT、AST、Cr和BUN水平均显著高于假手术组（均P < 0.001）。CLP + 2-AP组的ALT和AST水平显著低于CLP组（分别为27.88、11.33，均P < 0.001）；CLP + 2-AP组的Cr和BUN水平显著低于CLP组（分别为11.02、7.15，均P < 0.001）。与假手术组相比，CLP组促炎（IL-1β和TNF-α）和抗炎（IL-10）细胞因子水平显著升高（均P < 0.001）；CLP + 2-AP组的IL-1β和IL-10水平显著降低（均P < 0.001），但CLP + 2-AP组的TNF-α水平降低不显著（P = 0.33）。假手术组7天生存率为100%，CLP组为13.3%，2-AP组为86.7%，CLP + 2-AP组为20.0%。抑制PKR激活可轻微改善CLP模型小鼠7天生存率趋势（Mantel Cox检验分析，χ² = 0.0012，P = 0.97）。在脓毒症小鼠模型中，抑制PKR活性可降低血浆炎症因子表达，减少血液和腹腔细菌载量，保护器官功能，提示抑制PKR活性在脓毒症治疗中具有潜在应用价值。
