食用酒对脑组织Ngb、Hif-1α和Na^+,K^+-ATPase活力影响及其死亡机制

Effects of ethanol on Ngb、Hif-1α and Na^+,K^+-ATPase activity of brain and mechanism of death

目的探讨商品食用白酒灌胃的大鼠脑组织脑红蛋白(Ngb)、缺氧诱导因子1α(Hif-1α)和钠钾三磷酸腺苷酶(Na+,K+-ATPase)的活力变化,探讨急性大剂量给酒及其死亡机制。方法SD雄性大鼠56只随机分急性给酒死亡组、急性灌酒组、灌水对照组、慢性灌酒组。急性灌酒组一次性灌胃食用白酒(北京红星二锅头,体积分数为56%)15ml/kg,再分酒后0.5、2、4、6和12 h时间点组;急性给酒死亡组,在急性灌酒组剂量基础上,再额外腹腔注射酒至死(剂量约5 ml/只);灌水对照组给予同剂量食用水灌胃;慢性灌酒组连续灌酒1个月,前2周予8 ml/(kg.次)、后2周12 ml(kg.次),2次/d间隔8 h。采用免疫组织化学技术检测脑组织Ngb和Hif-1α的表达,紫外可见分光光度计检测脑组织Na+,K+-ATPase活力。结果急性给酒死亡组Hif-1αIOD值高于急性灌酒组、慢性灌酒组和灌水对照组(P<0.05),Na+,K+-ATPase活力均低于其他各组(P<0.05);Ngb IOD值高于急性灌酒的各时间点组,与最高值的急性灌酒2~4 h组比较,差异无统计学意义(P>0.05),亦高于慢性灌酒组和灌水对照组(P<0.05)。结论急性大剂量给酒后脑组织Ngb、Hif-1α表达增强、Na+,K+-ATPase活力降低,与乙醇及其代谢产物影响脑组织氧和能量代谢有关,可能是乙醇中毒及其死亡的重要毒理学机制之一。

Objective By investigating the changes of Ngb, Hif-1α expression and Na＋ ,K＋ -ATPase activity of brain in acute and chronic garage to large dose wine rats to discuss the possible death mechanisms of acute alcoholism. Methods Fifty-six male SD rats were divided into four groups in random, acute gavage to wine group were intragastrically given edible wine （56% V/V） 15 ml/kg once and killed after 0.5,2,4,6,12 h. Acute gavage to wine death group were additionally injected edible wine to abdominal cavity until death（dosage about 5 ml a rat） . Chronic garage to wine group were given 8 ml/kg in the first 2 weeks and 12 ml/kg in last 2 weeks twice every day with 8 hours＇ intervals. Sham garage to wine group were intragastrically given pure water. The whole brain was sampled to observe the expression of Ngb and Hif-1α by immunohistochemistry and the activity of Na＋ , K＋ -ATPase by spectrophotometry. Results Compared with acute gavage to wine group （killed after 2 - 4 h）, chronic gavage to wine group and sham gavage to wine group, the Hif- 1α IOD of acute gavage to wine death group increased in rats＇ brain tissue（ P 〈 0.05）.And the Na＋ ,K＋-ATPase activity of acute garage to wine death group was lower than other 3 groups（ P 〈 0.05）. In acute garage to wine group （killed after 2 - 4 h）, the Ngb IOD was higher than the five groups and acute gavage to wine death group, but the changes were not significant（ P 〉 0.05）, while the Ngb IOD of acute gavage to wine death group was higher than those of chronic gavage to wine group and Sham garage to wine group（ P 〈 0.05）. Conclusion The changes of the expression of Ngb and Hif-1α were strengthened and the activity of Na＋, K＋ -ATPase was descended in the brain tissue after gavage to large dose wine, which suggested that the alcoholism of nerve centre was relation with the obstruction of oxygen supplication and energy metabolism of brain neuron and the sodium pump＇ s serious malfunction. It could be one of mechanisms of alcoholism death.