吸入法建立急性一氧化碳中毒迟发型脑病大鼠模型

A model of delayed encephalopathy acute carbon monoxide poisoning established with inhalation method

目的建立符合临床特点的大鼠急性一氧化碳中毒迟发型脑病(DEACMP)模型。方法采用随机数字表法将认知功能筛查合格的雄性Wistar大鼠196只分为实验组和对照组,实验组164只,对照组32只。对照组吸入空气,实验组吸入一氧化碳(CO)气体,根据吸入CO浓度不同分为三组,分别为1000、3000、4000 ppm组,每组存活32只,连续吸入CO 40 min保持中毒浓度稳定;各组大鼠分别在中毒后7、14、21和28 d处死大鼠取材;采用分光光度计分析碳氧血红蛋白(COHb)水平;采用水迷宫训练评估各组大鼠中毒后认知功能的变化情况,采用HE染色方法观察大鼠海马细胞受损情况,明确建立可靠的大鼠DEACMP模型的方法。结果吸入CO后大鼠出现明显中毒表现,COHb水平与吸入CO浓度大小呈正相关(r = 0.748,P < 0.05),1000、3000、4000 ppm组及对照组大鼠死亡率分别为0%、40.74%、58.97%、0%,各组大鼠DEACMP发生率分别为6.25%、28.13%、31.25%、0%,在中毒后21、28 d大鼠DEACMP高发,选择死亡率低、发病率高、并结合大鼠饲养周期,明确大鼠吸入CO浓度3000 ppm,中毒后21 d可以建立DEACMP大鼠模型。结论大鼠吸入CO浓度3000 ppm 40 min,21 d后可以建立可靠DEACMP模型,为DEACMP的预防和治疗带来新的希望。

Objective To establish a model of delayed encephalopathy acute carbon monoxide poisoning (DEACMP). Methods A total of 196 intelligence-qualified rats were divided into experimental group and control group by random number table method, with 164 rats in the experimental group and 32 rats in the control group. The experimental group used an acute static inhalation of carbon monoxide (CO) to establish a poisoned rat model. According to different concentrations of CO poisoning, they were randomly divided into 1000, 3000, 4000 ppm group, with 32 rats alive in each group. Each group continued to inhale for 40 minutes. Rats in each group were sacrificed at 7, 14, 21 and 28 d after poisoning. Spectrophotometer method was used to dynamically monitor the changes of carboxyhemoglobin (COHb) content in each group. Morris water maze test was used to evaluate cognitive function. HE staining was used to observe the cell damage in hippocampus. The best poisoning concentration and time for establishing a DEACMP rat model was established. Results After poisoned by CO, the rats showed typical symptoms of poisoning. There was a significant positive correlation between COHb content and CO concentration (r = 0.748, P < 0.05). The mortality rates of the 1000, 3000, 4000 ppm group and the control group were 0%, 40.74%, 58.97% and 0%, respectively. The incidence of DEACMP in each group was 0%, 6.25%, 28.13%, 31.25%, and 0%, and the 21 d and 28 d after being poisoned were high DEACMP time points. Based on the low mortality rate and high incidence rate, the CO poisoning concentration of 3000 ppm and the time of 21d were the best method to establish a DEACMP rat model. Conclusion A DEACMP rat model can be established with a static inhalation concentration of 3000 ppm CO for 40 minutes, and models can be successfully established 21 days after poisoning.