Pavshintsev Vsevolod V, Mitkin Nikita A, Frolova Olga Y, Kushnir Ekaterina A, Averina Olga A, Lovat Maxim L
Faculty of Biology, Lomonosov Moscow State University, Leninskye Gory 1, 119234 Moscow, Russia.
Laboratory of Intracellular Signaling in Health and Disease, Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Vavilov Str. 32, 119991 Moscow, Russia.
Physiol Behav. 2017 Oct 1;179:458-466. doi: 10.1016/j.physbeh.2017.07.022. Epub 2017 Jul 20.
Alcohol dehydrogenases (ADH) are key enzymes of ethanol metabolism that mediate its oxidation to acetaldehyde. ADHs are also able to oxidize some types of neurotransmitters such as dopamine, serotonin and norepinephrine. Increased level of ADHs activity, induced by chronic alcohol consumption, is presumably associated with disturbed neurotransmitters metabolism that leads to stable alcohol craving. As earlier reported, intraperitoneal administration of 4-methilpirasole (non-specific inhibitor of ADHs) has shown to provide a short-term anti-alcoholic effect, but individual roles of ADH isoforms in this process were still unclear. The aim of this work was to study the roles of brain and serum ADH isoforms in alcohol consumption and neurotransmitter metabolism in the rats. In the study we used specific-pathogen-free (SPF) Wistar rats chronically alcoholized with 15% ethanol. 4-methilpirasole intranasal administration in small doses led to local inhibition of ADH III activity in the brain estimated by spectrophotometric assay. It correlated with dose-dependent reduction of dopamine concentration and increased level of its metabolic products in the brain but did not influence alcohol consumption. These data allowed us to propose an important role of brain ADHs (predominantly ADH III) in metabolism of dopamine in chronically alcoholized rats but not in regulation of alcohol consumption. To evaluate the role of serum ADH isoforms we immunized the rats with recombinant horse ADH that led to production of high levels of cross-reactive anti-ADH antibodies verified by ELISA assay. Immunization led to 30% decrease in alcohol consumption and recovery of general behavioral parameters such as motor activity, anxiety and depression level. At the same time active immunization did not cause any impairments in animal blood composition. We can conclude that immunization against ADHs appeared to be a safe way to decrease alcohol consumption that could be possibly associated with neurotransmitters metabolism correction.
乙醇脱氢酶（ADH）是乙醇代谢的关键酶，介导乙醇氧化为乙醛。ADH还能够氧化某些类型的神经递质，如多巴胺、血清素和去甲肾上腺素。长期饮酒诱导的ADH活性水平升高，可能与神经递质代谢紊乱有关，进而导致对酒精的稳定渴望。如先前报道，腹腔注射4-甲基吡唑（ADH的非特异性抑制剂）已显示出短期抗酒精作用，但ADH同工型在这一过程中的个体作用仍不清楚。这项工作的目的是研究大脑和血清ADH同工型在大鼠酒精摄入和神经递质代谢中的作用。在研究中，我们使用了用15%乙醇长期酒精化的无特定病原体（SPF）Wistar大鼠。小剂量鼻内给予4-甲基吡唑导致通过分光光度法测定的大脑中ADH III活性的局部抑制。这与大脑中多巴胺浓度的剂量依赖性降低及其代谢产物水平的升高相关，但不影响酒精摄入。这些数据使我们能够提出大脑ADH（主要是ADH III）在长期酒精化大鼠多巴胺代谢中起重要作用，但在酒精摄入调节中不起作用。为了评估血清ADH同工型的作用，我们用重组马ADH免疫大鼠，导致产生高水平的交叉反应抗ADH抗体，通过ELISA测定法验证。免疫导致酒精摄入量减少30%，并恢复了一般行为参数，如运动活动、焦虑和抑郁水平。同时，主动免疫未对动物血液成分造成任何损害。我们可以得出结论，针对ADH的免疫似乎是一种安全的减少酒精摄入的方法，这可能与神经递质代谢的纠正有关。
