一氧化碳-血红蛋白携氧载体对小鼠脓毒性脑病的作用及其机制  

Carbon monoxide-hemoglobin oxygen carrier improves mouse sepsis-associated encephalopathy by reducing inflammatory and oxidative stress in the hippocampus

作　　者：陈磊[1] 钱彬[1] 林芩[1] Chen Lei;Qian Bin;Lin Qin(Department of Anesthesiology,People’s Hospital Affiliated to Fujian University of Traditional Chinese Medicine,Fuzhou 350004,China)

机构地区：[1]福建中医药大学附属人民医院麻醉科,福州350004

出　　处：《创伤与急诊电子杂志》2023年第1期10-16,共7页Journal of Trauma and Emergency(Electronic Version)

基　　金：福建省中青年教师教育科研项目(JAT210159)。

摘　　要：目的探讨一氧化碳-血红蛋白携氧载体(carbon monoxide-hemoglobin based oxygen carrier,CO-HBOC)对脓毒性脑病(sepsis-associated encephalopathy,SAE)的作用及其机制。方法选取SPF级雄性C57BL/6小鼠136只,采用随机数字表随机分为4组:Sham组(假手术处理,术后1h尾静脉注射0.9%氯化钠溶液)、CLP组(建立脓毒症模型,术后1h尾静脉注射0.9%氯化钠溶液)、O2-HBOC组(建立脓毒症模型,术后1h尾静脉注射0.3 Hb/kg O2-HBOC)和CO-HBOC组(建立脓毒症模型,术后1h尾静脉注射0.3 Hb/kg CO-HBOC)。采用盲肠结扎穿刺法建立脓毒症小鼠模型,Morris水迷宫实验观察认知功能。术后24h取脑组织切片采用原位末端标记法(TdT-mediated dUTP nick end labeling,TUNEL)/神经元核(neuronal nuclei,NeuN)/4,6-二氨基-2-苯基吲哚(4,6-diamino-2-phenyl indole,DAPI)荧光染色观察海马凋亡神经元比例;检测海马组织肿瘤坏死因子α(tumor necrosis factorα,TNF-α)、白介素-6(interleukin-6,IL-6)、白介素-1β(interleukin-1β,IL-1β)水平,以及线粒体活性氧(mitochondrial reactive oxygen species,mtROS)含量、丙二醛(malonic dialdehyde,MDA)水平。结果与CLP组相比,CO-HBOC组小鼠水迷宫训练期第3、4天逃避潜伏期减少(t值分别为2.559、2.967,P<0.05),测试期目标象限停留时间增加[(18.4±6.3)s比(8.3±5.5)s,t=3.801,P<0.001],平台穿越次数增加[(3.6±1.8)次比(1.8±1.2)次,t=2.774,P=0.009];COHBOC组海马凋亡神经元比例减少[(16.8±6.2)%比(31.6±8.4)%,t=4.184,P<0.001];CO-HBOC组小鼠海马TNF-α水平降低[(110.3±21.2)ng/L比(205.4±35.5)ng/L,t=6.286,P<0.001],IL-6水平降低[(67.1±14.3)ng/L比(100.6±16.7)ng/L,t=4.218,P<0.001],IL-1β水平降低[(2.1±0.3)ng/L比(3.4±0.6)ng/L,t=5.456,P<0.001],mtROS水平降低[(140.1±24.3)%比(239.8±33.4)%,t=7.482,P<0.001],MDA含量减少[(14.0±3.5)nmol/mg蛋白比(24.6±4.0)nmol/mg蛋白,t=4.699,P<0.001]。结论CO-HBOC可能通过减轻海马神经炎症与氧化应激反应,降低海马神经元凋亡水平,改善SAE�Objective To investigate the effect of carbon monoxide-hemoglobin based oxygen carrier(CO-HBOC)on sepsis-associated encephalopathy(SAE)in mice.Method The SPF male C57BL/6 mice were randomly divided into 4 groups:sham group,CLP group,O2-HBOC group and CO-HBOC group.The animal model of sepsis was established by cecal ligation and puncture(CLP).The Morris water maze(MWM)test was used to observe the cognitive function of mice.We calculated the proportion of apoptotic neurons in the hippocampus by TdT-mediated dUTP nick end labeling(TUNEL)staining at 24 h postoperatively.The hippocampal levels of the inflammatory factors including tumor necrosis factorα(TNF-α),interleukin-6(IL-6),interleukin-1β(IL-1β)and the levels of mitochondrial reactive oxygen species(ROS)and malonic dialdehyde(MDA)were also evaluated at 24 h postoperatively.Result Compared with the CLP group,the escape latency of the CO-HBOC group on the 3rd and 4th days of the MWM training period decreased;the target quadrant residence time[(18.4±6.3)s vs(8.3±5.5)s],and the number of platform crossings increased[(3.6±1.8)vs(1.8±1.2)].The proportion of apoptotic neurons in the hippocampus of hippocampal decreased in the CO-HBOC group[(16.8±6.2)%vs(31.6±8.4)%].CO-HBOC treatment also decreased the levels of hippocampal TNF-α[(110.3±21.2)ng/L vs(205.4±35.5)ng/L],IL-6[(67.1±14.3)ng/L vs(100.6±16.7)ng/L],IL-1β[(2.1±0.3)ng/L vs(3.4±0.6)ng/L],mitochondrial ROS[(140.1±24.3)%vs(239.8±33.4)%]and MDA[(14.0±3.5)nmol/mg protein vs(24.6±4.0)nmol/mg protein].Conclusion CO-HBOC may reduce the level of apoptosis of hippocampal neurons and improve SAE in mice by reducing the neuroinflammation and oxidative stress in the hippocampus.

关 键 词：脓毒性脑病 认知功能 神经炎症 一氧化碳-血红蛋白携氧载体 氧化应激 

分 类 号：R459.7[医药卫生—急诊医学] R747.9[医药卫生—治疗学]