一氧化碳中毒迟发性脑病小鼠脑HO-1表达变化及其与细胞凋亡的关系

Changes of heme oxygenase-1 and apoptosis as well as the relationship between them following carbon monoxide poisoning delayed encephalopathy in mice

目的研究一氧化碳(carbon monoxide,CO)中毒后不同时间点小鼠脑内血红素加氧酶-1(heme oxygenase-1 HO-1)的表达变化;检测CO中毒后不同时间点小鼠脑内神经细胞凋亡及相关基因表达的情况。探讨血红素氧化酶-1与脑细胞凋亡之间的变化关系。方法使用腹腔注射CO的方法制备CO中毒迟发性脑病(Delayed encephalopathy after acute carbon monoxide poisoning,DEACMP)的模型,应用Western blotting法观察HO-1蛋白水平的表达,应用流式细胞仪法检测凋亡细胞,采用SABC免疫组化法观察Bcl-2、Bax在蛋白水平的表达。结果 Western blotting法HO-1蛋白表达情况:空气对照组表达较少,CO中毒组1 d表达增加(1.14±0.43,P<0.01),3 d达高峰(2.91±1.07,P<0.01),21 d仍高于对照组(P<0.01)。细胞凋亡情况:空气对照组各时间点细胞凋亡百分数较少,CO中毒组12 h凋亡细胞百分数数增加(21.95±24,P<0.01),5 d达高峰(34.27±4.19,P<0.01),21 d时(9.25±1.16)仍高于对照组。凋亡相关蛋白表达:Bax在中毒后表达增加,3 d达高峰(34.5±0.58,P<0.01),Bcl-2在3 d减少至最低(12.5±3.69,P<0.01),Bax/Bcl-2的比值于3 d时达最大。结论 CO中毒迟发性脑病小鼠海马区HO-1蛋白表达明显增高,凋亡细胞明显增多且与HO-1的表达变化一致。HO-1表达增高可能参与了CO中毒所致脑损伤的发病机制。

Objective To study the expression of the heme oxygenase- 1 in the brain of mice at different time points after carbon monoxide poisoning. To detect neuron apoptosis and related gene expression in the brain of mice at different time points after carbon monoxide poisoning. To investigate the relationship between changes in heme oxygenase- 1 and apoptosis of brain cells. Methods Carbon monoxide poisoning delayed encephalopathy model by using intraperitoneal injection of carbon monoxide. Application The expression of HO- 1 protein levels was observed By Western blotting analysis. Flow cytometry was used to detect apoptotic cells. Application immunohistochemistry SABC observed Bcl-2 and Bax expression at the protein level. Results Expression of HO- 1 protein is less in the air Control group by Western blotting; expression increased at ld （1.14±0.43） （P〈0.01）, reaching a peak at 3d （2.91±1.07） （P〈0.01） in CO poisoning group; Expression of HO-1 protein in poisoning group was still higher than in that in the control group at 21d （P〈0.01）. Apoptosis： the percentage of apoptotic cells increased in CO poisoning 12h group （21.95±24） （P〈 0.01）, reaching the peak at 5d （34.27±4.19） （P〈 0.01）. The percentage of apoptotic cells was still high at 21d （9.25±1.16）. Protein related： Bax expression increased after the poisoning, reaching the peak at 3d （34.5±0.58） （P〈0.01）. Bcl-2 expression reduced to a minimum at 3d （12.5±3.69） （P〈0.01）. The ratio of Bax/Bcl-2 reached maximum at 3d. Conclusion Expression of heme oxygenase- 1 protein and Apoptotic cells were significantly increased in the hippocampus after delayed encephalopathy in mouse of carbon monoxide poisoning. The changes of apoptotic cells were consistent with heme oxygenase- 1 protein expression. The upregnlated expression of heme oxygenase- 1 protein plays a crucial role in the pathogenesis of delayed encephalopathy after CO poisoning.