IL-18抑制剂对脓毒症并发血小板减少小鼠的保护作用

Protective effects of IL-18 inhibitors on sepsis associated thrombocytopenia in mice

目的:探究白细胞介素(IL)-18抑制剂(IL-18Bp)对脓毒症并发血小板减少小鼠的保护作用。方法:将30只C57BL/6雄性小鼠依次编号,按照随机数字表法分为3组:对照组、脂多糖(LPS)注射模型组(LPS组)、LPS加IL-18Bp干预组(LPS/IL-18Bp组)。其中LPS组和LPS/IL-18Bp组给予单次腹腔注射30 mg/kg LPS制备脓毒症并发血小板减少模型,对照组给予等体积生理盐水;LPS/IL-18Bp组在注射LPS后给予单次腹腔注射50μg/kg IL-18Bp,其余各组给予等体积生理盐水。观察各组72 h生存率,并于LPS注射后24 h时收集血液样本和脾脏组织,全血细胞分析仪检测血小板计数,ELISA法检测各组富血小板血浆中血小板活化标志物sCD40L以及IL-18和IL-6、IL-10、肿瘤坏死因子(TNF)-α的表达情况;HE染色观察并分析各组脾脏的病理变化。结果:LPS/IL-18Bp组各时间点的生存率均高于LPS组(χ^(2)=0.63,P<0.05);LPS注射后24 h时LPS/IL-18Bp组血小板计数显著高于LPS组(t=8.37,P=0.001);LPS/IL-18Bp组sCD40L表达量较LPS组低(t=4.34,P=0.012);LPS/IL-18Bp组小鼠富血小板血浆中IL-18、IL-6、IL-10和TNF-α的表达均较LPS组显著降低(t=8.18、10.95、5.34、10.68,均P<0.05);脾脏病理分析显示,LPS/IL-18Bp组脾脏炎症细胞较LPS组明显减少。结论:IL-18Bp能够缓解LPS诱导小鼠血小板减少、血小板活化和炎症反应,并提高LPS诱导血小板减少小鼠模型的生存率,对LPS诱导血小板减少模型小鼠具有一定的保护作用。

Objective:To investigate the protective effect of interleukinc(IL)-18 inhibitor(IL-18Bp)on mice with sepsis complicated with thrombocytopenia(TCP).Methods:Thirty C57BL/6 male mice were numbered sequentially,according to the random number table method,they were divided into three groups:control group,lipopolysaccharide(LPS)injection model group(LPS group),and IL-18Bp intervention group(LPS/IL-18Bp group).The model of sepsis mice with thrombocytopenia was prepared by single intraperitoneal injection of 30 mg/kg LPS in the LPS group and the LPS/IL-18Bp group,and the control group was given constant volume of normal saline.Mice in the LPS/IL-18Bp group were given a single intraperitoneal injection of 50μg/kg IL-18Bp after LPS injection,and the other groups were given the same volume of normal saline.The survival rate of mice in each group after 72 hours were observed.Blood samples and spleen tissues were collected 24 hours after LPS injection.The platelet count was measured by a whole blood cell analyzer.The expression of platelet activation marker sCD40L,IL-18 and inflammatory factors such as IL-6,IL-10 and tumor necrosis factor(TNF)-αin platelet-rich plasma was detected by enzyme-linked immunosorbent assay(ELISA).The pathological changes of spleen in each group were observed and analyzed by hematoxylin-eosin staining(HE).Results:The survival rate of mice in the IL-18Bp intervention group was higher than that in the LPS group at all time points(χ^(2)=0.63,P<0.05).At 24 hours after LPS injection,the platelet count of IL-18Bp intervention group was significantly higher than that of LPS group(t=8.37,P=0.001).The expression of sCD40L in IL-18Bp intervention group was lower than that in LPS group(t=4.34,P=0.012).The expressions of IL-18,IL-6,IL-10 and TNF-αin platelet-rich plasma of mice in IL-18Bp intervention group were significantly lower than those in LPS group(t=8.18,10.95,5.34,10.68,all P<0.05).The pathological analysis of spleen showed that the inflammatory cells in the spleen of mice of the IL-18Bp intervention group were significantly reduced compared with the LPS group.Conclusion:IL-18Bp can alleviate thrombocytopenia,platelet activation and inflammation in TCP mice induced by LPS,and improve the survival rate of TCP mice induced by LPS,and has a certain protective effect on the model of TCP mice induced by LPS.