IL-18结合蛋白在创伤性颅脑损伤大鼠认知功能障碍中的作用及其机制.pdf

解放军医学杂志2023年7月28日第48卷第7期776中图分类号R651.1文献标志码ADOI10.11855/j.issn.0577-7402.1618.2023.0103声明本文所有作者声明无利益冲突引用本文李妍,齐曼曼,缴宝杰,等.IL-18结合蛋白在创伤性颅脑损伤大鼠认知功能障碍中的作用及其机制J.解放军医学杂志,2023,48(7):776-783.收稿日期2022-07-28录用日期2022-09-17上线日期2023-01-03IL-18结合蛋白在创伤性颅脑损伤大鼠认知功能障碍中的作用及其机制李妍1，齐曼曼1，缴宝杰1，李春雷1，王春雷2，常玉林1*1沧州市中心医院麻醉科，河北沧州061000；2沧州市中心医院运动医学科，河北沧州061000基础研究基金项目河北省医学科学研究课题(20220360)作者简介李妍，医学硕士，主治医师，主要从事围手术期神经功能损伤方面的研究通信作者常玉林，E-mail：论著摘要目的探究IL-18结合蛋白(IL-18BP)在创伤性脑损伤(TBI)大鼠认知功能障碍中的作用及其机制。方法120只健康雄性SD大鼠按随机数字表法分为假手术组、TBI组、TBI+IL-18BP组(经尾静脉注射IL-18BP 1.5 mg/kg)与TBI+IL-18BP+ADU-S100组(经尾静脉注射IL-18BP 1.5 mg/kg，经腹腔注射ADU-S100 20 mg/kg)，每组30只。应用自由落体法建立TBI模型，造模后30 d行水迷宫实验检测大鼠认知功能，ELISA法检测大鼠血清IL-18、IL-18BP浓度，免疫荧光染色检测海马区星形胶质细胞GFAP、IL-18和cleaved-caspase-3荧光强度，Western blotting检测大鼠海马区干扰素基因刺激蛋白(STING)、磷酸化TANK结合激酶1(p-TBK1)、TBK1、磷酸化干扰素调节因子3(p-IRF3)、IRF3蛋白相对表达量。结果与假手术组比较，TBI组、TBI+IL-18BP组和TBI+IL-18BP+ADU-S100组大鼠逃逸潜伏期明显延长，穿越平台次数减少，目标象限活动时间百分比降低(P0.0001)，血清IL-18、IL-18BP浓度升高，海马区IL-18和cleaved-caspase-3荧光强度明显增强，STING、p-TBK1、p-IRF3表达明显上调(P0.05)；与TBI组比较，TBI+IL-18BP组大鼠逃逸潜伏期缩短，穿越平台次数增多，目标象限活动时间百分比增高(P0.0001)，血清IL-18浓度降低，IL-18BP浓度升高，海马区IL-18和cleaved-caspase-3荧光强度明显减弱，STING、p-TBK1、p-IRF3表达明显下调(P0.05)；与TBI+IL-18BP组比较，TBI+IL-18BP+ADU-S100组大鼠逃逸潜伏期延长，穿越平台次数减少，目标象限活动时间百分比降低(P0.0001)，血清IL-18浓度升高，IL-18BP浓度降低，海马区IL-18和cleaved-caspase-3荧光强度增强，STING、p-TBK1、p-IRF3表达明显上调(P0.05)。结论IL-18BP可改善TBI大鼠的认知功能，其机制可能与抑制星形胶质细胞凋亡及抑制STING/TBK1/IRF3信号通路激活有关。关键词IL-18结合蛋白；认知功能；星形胶质细胞；细胞凋亡；干扰素基因刺激蛋白Effect and mechanism of IL-18BP in relieving cognitive dysfunction of rats with traumatic brain injuryLi Yan1,Qi Man-Man1,Jiao Bao-Jie1,Li Chun-Lei1,Wang Chun-Lei2,Chang Yu-Lin1*1Department of Anesthesiology,2Department of Sport Medicine,Cangzhou Central Hospital,Cangzhou,Hebei 061000,China*Corresponding author,E-mail:This work was supported by the Medical Science Research Project of Hebei Province(20220360)AbstractObjectiveTo explore the role of interleukin-18 binding protein(IL-18BP)in alleviating cognitive dysfunction of traumatic brain injury(TBI)rats and the related mechanisms.MethodsA total of 120 adult male SD rats were randomly divided into 4 groups(30 each):sham group,TBI group,TBI+IL-18BP group(administered 1.5 mg/kg IL-18BP by tail vein),and TBI+IL-18BP+ADU-S100 group(administered 1.5 mg/kg IL-18BP by tail vein,and 20 mg/kg ADU-S100 by intraperitoneal injection).The rat model of TBI was established using the free falling body method.At 30 d after modeling,cognitive function of rats was measured by Morris water maze test.The serum levels of IL-18 and IL-18BP were detected by ELISA.The integrated fluorescence intensity Med J Chin PLA,Vol.48,No.7,July 28,2023777of GFAP,IL-18 and cleaved-caspase-3 in hippocampus were detected by immunofluorescence.The relative expression levels of stimulator of interferon genes(STING),phosphorylated TANK-binding kinase 1(p-TBK1),TBK1,phosphorylated interferon regulating factor 3(p-IRF3),and IRF3 in hippocampus were detected using Western blotting.ResultsCompared with sham group,rats in TBI,TBI+IL-18BP and TBI+IL-18BP+ADU-S100 groups had longer escape latency and decreased times of crossing the platform and reduced time of acting in the targeted quadrant(P0.0001),with increased concentration of IL-18 and IL-18BP in the blood,enhanced fluorescence intensity of IL-18 and cleaved-caspase-3,up-regulated expression levels of STING,p-TBK1 and p-IRF3 in the hippocampus(P0.05);Compared with TBI group,the escape latency was shortened,the times of crossing platform and the percentage of target quadrant activity time were increased in TBI+IL-18BP group(P0.001),the concentration of serum IL-18 decreased while of serum IL-18BP increased,and the fluorescence intensity of IL-18 and cleaved-caspase-3,the expressions of STING,p-TBK1 and p-IRF3 in the hippocampus were down-regulated significantly in TBI+IL-18BP group(P0.05);Compared with TBI+IL-18BP group,the latency was significantly prolonged,the platform crossing times remarkably reduced,the time spent in the target quadrant was considerably shortened,the concentration of serum IL-18 increased while of IL-18BP decreased,the fluorescence intensity of IL-18 and cleaved-caspase-3,the expression levels of STING,p-TBK1 and p-IRF3 in hippocampus were increased in TBI+IL-18BP+ADU-S100 group(P0.05).ConclusionIL-18BP can improve the cognitive function of TBI rats to some extent,and its mechanism may be related to the inhibition of astrocytes apoptosis and STING/TBK1/IRF3 signaling pathway.Key wordsinterleukin-18 binding protein;cognitive function;astrocyte;apoptosis;stimulator of interferon genes创伤性脑损伤(traumatic brain injury，TBI)为暴力打击等因素导致的局灶性损伤、颅内出血、脑水肿、脑缺血等，是全球范围内造成死亡和残疾的主要原因之一1。创伤幸存者常伴有不同程度的、持续时间较长的认知功能障碍，主要表现为学习、记忆能力下降2-3。术后认知功能障碍(post-operative cognitive dysfunction，POCD)是指麻醉手术后患者存在记忆力、抽象思维、定向力障碍，并伴有人格、社交能力的改变4，具有造成患者康复延迟、并发症增多、住院时间延长及医疗费用增加的风险。因此，寻找POCD的防治方法、改善术后认知康复尤为重要。TBI可引发原发性损伤和继发性损伤，其中继发性损伤由线粒体功能障碍、氧化应激、脂质过氧化、神经炎症、轴突变性和细胞凋亡等因素导致5-6。星形胶质细胞在TBI的发病机制中有利有弊，如促进或限制神经发生和突触发生，加速或抑制神经炎症，以及通过多种生物活性因子破坏或修复血脑屏障等7-9。在脑外伤患者和实验动物模型中，星形胶