C-EPO改善创伤性脑损伤合并海水浸泡性体温过低症大鼠的实验研究  

Carbamylated erythropoietin ameliorating hypothermia in rats with traumatic brain injury combined with seawater immersion

作　　者：吴再新 张灏 潘振洪 陈宏颉 郑兆聪 魏梁锋 叶丹[3] 王守森 Wu Zaixin;Zhang Hao;Pan Zhenhong;Chen Hongjie;Zheng Zhaocong;Wei Liangfeng;Ye Dan;Wang Shousen(Department of Military Medical Service,No.900 Hospital of PLA,Fuzhou 350025,China;Department of Neurosurgery,Fuzong Clinical Medical College of Fujian Medical University(No.900 Hospital of PLA),Fuzhou 350025,China;Department of Neurosurgery,Fuzong Teaching Hospital of Fujian University of Traditional Chinese Medicine(No.900 Hospital of PLA),Fuzhou 350025,China)

机构地区：[1]联勤保障部队第九〇〇医院卫勤部,福州350025 [2]福建医科大学福总临床医学院(联勤保障部队第九〇〇医院)神经外科,福州350025 [3]福建中医药大学福总教学医院(联勤保障部队第九〇〇医院)神经外科,福州350025

出　　处：《中华航海医学与高气压医学杂志》2024年第2期163-168,共6页Chinese Journal of Nautical Medicine and Hyperbaric Medicine

基　　金：全军后勤科技重大项目子课题(AHJ14J001);中国人民解放军联勤保障部队第九〇〇医院院内课题(2022ZL01,2022ZL03)

摘　　要：目的评价氨甲酰化促红细胞生成素(C-EPO)对创伤性脑损伤合并海水浸泡性体温过低症(TBI-SIH)大鼠的影响及其与NOD样受体热蛋白结构域相关蛋白3(NLRP3)炎症小体的关系。方法36只SD大鼠按照随机数字表法分为sham组(n=12)、TBI+SIH组(n=12)和C-EPO治疗组(n=12),采用受控皮质撞击颅脑致伤法+恒温水浴浸泡法建立TBI合并SIH大鼠模型,C-EPO治疗组每24 h腹腔注射50μg/kg的C-EPO。于模型制备后48 h进行改良神经功能缺损评分(mNSS)和旷场实验,行为学测定结束后,处死大鼠,测定脑组织含水量及伊文思蓝(EB)含量,采用ELISA法检测损伤区域脑组织IL-1β、IL-18及caspase-1含量,采用qPCR实验检测损伤区脑组织NLRP3 mRNA、caspase-1 mRNA和IL-18 mRNA表达水平。结果与sham组比较,TBI+SIH组大鼠mNSS升高[(9.9±0.8)分],运动总路程[(26743±2034)mm]和中央区域停留时间[(7.7±1.5)s]减少,脑组织含水量[(82.16±3.25)%]及EB含量[(33.49±5.28)μg/g]升高,损伤区脑组织IL-1β[(476.0±34.7)pg/mg]、IL-18[(1501.0±113.2)pg/mg]和caspase-1[(1873.0±156.0)pg/mg]含量升高,NLRP3 mRNA(1.48±0.19)、caspase-1 mRNA(1.90±0.49)和IL-18 mRNA(2.92±0.63)表达上调(P<0.05);与TBI+SIH组比较,C-EPO治疗组大鼠mNSS[(5.2±0.7)分]降低,运动总路程[(45901±3425)mm]和中央区域停留时间[(21.0±3.5)s]增加,脑组织含水量[(80.79±3.40)%]及EB含量[(19.23±3.76)μg/g]降低,损伤区脑组织IL-1β[(412.0±22.0)pg/mg]、IL-18[(660.0±45.8)pg/mg]和caspase-1[(1123.0±121.9)pg/mg]含量降低,NLRP3 mRNA(0.96±0.11)、caspase-1 mRNA(0.93±0.30)和IL-18 mRNA(1.28±0.35)表达下调(P<0.05)。与sham组比较,TBI+SIH组大鼠出现明显的脑水肿及脑出血;与TBI+SIH组相比,C-EPO治疗组大鼠脑水肿反应减轻,脑组织结构更清晰,脑出血程度也明显减轻。结论C-EPO可减轻TBI合并SIH大鼠脑损伤,其机制可能与抑制NLRP3炎症小体激活有关。Objective To evaluate the effect of carbamylated erythropoietin(C-EPO)on hypothermia in rats with traumatic brain injury combined with seawater immersion(TBI-SIH),and its relationship with NOD-like receptor thermal protein domain associated protein 3(NLRP3)inflammasomes.Methods A total of 36 Sprague-Dawley(SD)rats were randomly divided into three groups:sham group(n=12),TBI+SIH group(n=12),and C-EPO treatment group(n=12).The controlled cortical impact(CCI)TBI injury method and constant temperature water bath immersion method were employed to establish a TBI+SIH rat model.C-EPO was administered at a dose of 50μg/kg every 24 hours.Forty-eight hours after modeling,the modified neurological severity score(mNSS)and open field test were conducted.The ELISA was employed to test the levels of IL-1β,IL-18,and Caspase-1 within the injury region,while quantitative polymerase chain reaction(qPCR)was utilized to assess the expression levels of NLRP3 mRNA,caspase-1 mRNA,and IL-18 mRNA.Results Compared with the sham group,the TBI+SIH group exhibited increased mNSS values[(9.9±0.8)points],and decreased total movement distance[(26743±2034)mm]and stay time in central area[(7.7±1.5)s]in the open field test,elevated brain water content[(82.16±3.25)%]and EB content[(33.49±5.28)μg/g],and increased levels of IL-1β[(476.0±34.7)pg/mg],IL-18[(1501.0±113.2)pg/mg],and caspase-1[(1873.0±156.0)pg/mg]in the injury area,and significantly upregulated expressions of NLRP3 mRNA(1.48±0.19),Caspase-1 mRNA(1.90±0.49),and IL-18 mRNA(2.92±0.63)(P<0.05).Compared with the TBI+SIH group,the C-EPO treatment group showed reduced mNSS scores[(5.2±0.7)points],increased total movement distance[(45901±3425)mm]and stay time in central area[(21.0±3.5)s]in the open field test,decreased brain water content[(80.79±3.40)%]and EB content[(19.23±3.76)μg/g],reduced levels of IL-1β[(412.0±22.0)pg/mg],IL-18[(660.0±45.8)pg/mg],and caspase-1[(1123.0±121.9)pg/mg]in the injured cortex,and downregulated expressions of NLRP3 mRNA(0.96±0.11),Caspase-1 m

关 键 词：创伤性脑损伤 海水浸泡性体温过低症 氨甲酰化促红细胞生成素 NLR家族 热蛋白结构域相关蛋白3 

分 类 号：R651.15[医药卫生—外科学]