大鼠急性一氧化碳中毒迟发性脑病模型中干预剂叔丁基对苯二酚对Nrf2及HO-1表达的影响

Effect of tBHQ on expression of Nrf2 and HO-1 in delayed encephalopathy in rats after acute carbon monoxide poisoning

目的:初步探讨大鼠急性一氧化碳中毒迟发性脑病(DEACMP)模型中干预剂叔丁基对苯二酚(t BHQ)前后脑组织海马区核因子E2相关因子2(Nrf2)及其下游靶基因血红素加氧酶1(HO-1)的变化,从而进一步研究DEACMP的发病机制,同时为其靶向治疗的研究提供一定的实验基础。方法:将240只大鼠按随机数字表法分为一氧化碳中毒组(CO组)、空气对照组(AC组)、一氧化碳+3%乙醇组(EC组)、一氧化碳+t BHQ组(TC组),再将各组大鼠按染毒后1、3、7、14、21、28 d随机分为6个亚组,随后用Morris水迷宫试验观察大鼠行为学表现,免疫组织化学及蛋白免疫印迹法(Western blot)检测大鼠干预剂t BHQ前后Nrf2及HO-1的表达变化,TUNEL染色检测细胞凋亡情况。结果:CO组、EC组、TC组大鼠海马区Nrf2及HO-1表达均表现为染毒后第1天升高,3 d达高峰,随后逐渐下降的趋势,各时间点与AC组比较差异均有统计学意义,TC组与CO组比较Nrf2及HO-1表达增高,各亚组比较差异均有统计学意义。CO组、EC组、TC组大鼠与AC组比较凋亡细胞随时间明显增多,且均表现为增高-高峰(7~14 d)-降低的趋势,TC组与CO组比较凋亡细胞(7~14 d)减少,差异有统计学意义。结论:DEACMP的发生发展中Nrf2/ARE/HO-1通路起着重要作用,t BHQ特异性激活Nrf2通路达到早期保护作用,有望减少或减轻DEACMP。

Objective： Preliminary study on rats with delayed encephalopathy after acute carbon monoxide poisoning（ DEACMP） model intervention agent t BHQ before and after the nuclear factor E2-related factor 2（ Nrf2） and its downstream target genes of heme oxygenase 1（ HO-1） change in hippocampus associated,in order to further study the pathogenesis of DEACMP,at the same time for the targeted therapy of provide a certain experimental basis. Methods： One hundred and twenty rats were randomly divided into carbon monoxide poisoning group（ CO group）,air control group（ AC group）,carbon monoxide＋3% ethanol group（ EC group）,carbon monoxide＋t BHQ group（ group TC）,then the rats in exposure after 1 d,3 d,7 d,14 d,21 d,28 d,with the machine was divided into 6 sub groups,followed by the Morris water maze test to observe the behavior of rats,immunohistochemistry and protein Western blot method of chemical（ Western blot） detecting expression of Nrf2 and HO-1 mploying intervention agent t BHQ before and after,and then TUNEL staining was detected cell apoptosis. Results： CO group,EC group,TC group Nrf2 in hippocampus of rats and the expression of HO-1 were increased in the first day and reach a peak at the third day,then gradually decreased,and at each time point in AC group were statistically significant,TC group and CO group Nrf2 and HO-1 were increased in each sub group and the deffirences were statistically meaning. Comparison apoptotic cells in CO group,EC group,TC group with AC group rats increased significantly over time,and showed higher peak（ 7-14 d）-decreased. TC group compared with CO group,the apoptotic cells（ 7-14 d） decreased,the difference was statistically significant. Conclusion： The Nrf2/ARE/HO-1 pathway plays an important role in the development of DEACMP,and the t BHQ specific activation of the Nrf2 pathway achieves early protection and is expected to reduce or mitigate DEACMP.