摘要
目的:分析NF-κB/n NOS信号通路在急性CO中毒迟发性脑病(DEACMP)中的作用。方法:将昆明雄性小鼠随机分为空气组与CO中毒组,其中空气组和CO中毒组又分别分为1、3、7、14和21 d亚组。CO中毒组小鼠首次腹腔注射99. 9%CO气体100 ml/kg,随后3次按首次剂量的半量(50 ml/kg)作追加注射,间隔为4 h,空气组以同样的方法注射等量空气,立即观察其精神状态和行为变化;并通过改良双波长碳氧血红蛋白(COHB)定量法动态监测小鼠血中COHB浓度;使用旷场实验和筑巢实验进行行为学监测;并用ELSIA法测定脑组织内髓鞘碱性蛋白(MBP)含量,筛选出DEACMP模型小鼠。用ELISA法分别检测脑组织中NF-κB和n NOS蛋白,用紫外分光光度计法测定脑组织中一氧化氮(NO)含量。结果:与空气组相比,CO中毒组旷场实验垂直站立次数和进入旷场中心次数均明显减少(P <0. 01),CO中毒组筑巢得分明显下降(P <0. 05),CO中毒组小鼠MBP、NF-κB、n NOS蛋白水平及NO含量于第3、7、14和21 d均明显增高(P <0. 01)。结论:NF-κB/n NOS信号通路可能参与了DEACMP的发生和发展。
Objective: To analyze the role of NF-κB/n NOS signaling pathway in delayed encephalopathy( DEACMP)after acute carbon monoxide( CO) poisoning in mice. Methods: Kunming male mice were divided into air group and CO poisoning group. The air group and CO poisoning group mice were divided into 1,3,7,14 and 21 d subgroups,respectively. The CO poisoning group mice were injected intraperitoneally with 99. 9% CO gas 100 ml/kg in the first time,following injection 50 ml/kg CO three times and the interval time was 4 h each time. The air group mice were injected with equal amounts of air in the same manner. The blood carboxyhemoglobin( COHB) concentration via dual-wavelength COHB quantification; The behavior changes were observed in the open field test and the nesting test; and the protein level of myelin basic protein( MBP) of brain were monitored by ELISA. The levels of protein of NF-κB,n NOS were assayed by ELISA and the content of nitric oxide( NO) were detected by ultraviolet spectrophotometer. Results: In open field test,the number of vertical standing and the number of entering the center area were significantly reduced( P < 0. 01) in CO poisoning group compared with the air group. The nesting score was significantly decreased after the CO treatment in thenest test( P < 0. 05). The levels of MBP,NF-κB,n NOS and NO in the CO poisoning group were significantly increased on the 3,7,14 and 21 d compared with control group( P < 0. 01). Conclusion: NF-κB/n NOS signaling pathway may play a role in the pathology of DEACMP.
引文
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