Department of Immunology, College of Medicine, Institute of NeuroImmune Pharmacology, Florida International University, Miami, Florida 33199, USA.
Alcohol Clin Exp Res. 2009 Oct;33(10):1731-8. doi: 10.1111/j.1530-0277.2009.01010.x. Epub 2009 Jul 1.
Alcohol is the most widely abused substance and its chronic consumption causes neurobehavioral disorders. It has been shown that alcohol affects the function of immune cells. Dendritic cells (DC) serve as the first line of defense against infections and are known to accumulate neurotransmitters such as 5-hydroxytryptamine (5-HT). The enzyme monoamine oxidase-A (MAO-A) degrades 5-HT that is associated with clinical depression and other neurological disorders. 5-HT is selectively transported into neurons through the serotonin transporter (SERT), which is a member of the sodium- and chloride-dependent neurotransmitter transporter (SLC6) family. SERT also serves as a receptor for psychostimulant recreational drugs. It has been demonstrated that several drugs of abuse such as amphetamine and cocaine inhibit the SERT expression; however, the role of alcohol is yet to be elucidated. We hypothesize that alcohol can modulate SERT and MAO-A expression in DC, leading to reciprocal downregulation of 5-HT in extracellular medium.
Dendritic cells were treated with different concentrations (0.05% to 0.2%v/v) of alcohol for 24-72 hours and processed for SERT and MAO-A expression using Q-PCR and Western blots analysis. In addition, SERT function in DC treated with alcohol both in the presence and absence of imipramine, a SERT inhibitor was measured using 4-[4-(dimethylamino)styryl]-1-methylpyridinium iodide uptake assay. 5-HT levels in culture supernatant and intracellular 5-hydroxy indole acetic acid (5-HIAA) and cyclic AMP were also quantitated using ELISA.
Dendritic cells treated with 0.1% alcohol for 24 hours showed significant upregulation of SERT and MAO-A expression compared with untreated DC. We also observed that 0.1% alcohol enhanced the function of SERT and decreased extracellular 5-HT levels compared with untreated DC cultures, and this was associated with the elevation of intracellular 5-HIAA and cyclic AMP levels.
Our study suggests that alcohol upregulates SERT and MAO-A by elevating cyclic AMP, which may lead to decreased concentration of 5-HT in the extracellular medium. As 5-HT is a major neurotransmitter and an inflammatory mediator, its alcohol-mediated depletion may cause both neurological and immunological deregulation.
酒精是最广泛滥用的物质,其慢性消费会导致神经行为障碍。已经表明,酒精会影响免疫细胞的功能。树突状细胞(DC)作为对抗感染的第一道防线,已知会积累 5-羟色胺(5-HT)等神经递质。单胺氧化酶-A(MAO-A)降解与临床抑郁症和其他神经紊乱相关的 5-HT。5-HT 是通过血清素转运体(SERT)选择性地转运到神经元中的,SERT 是钠和氯离子依赖的神经递质转运体(SLC6)家族的成员。SERT 也作为精神兴奋剂娱乐药物的受体。已经证明,几种滥用药物,如安非他命和可卡因,抑制 SERT 的表达;然而,酒精的作用尚待阐明。我们假设酒精可以调节 DC 中的 SERT 和 MAO-A 表达,导致细胞外介质中 5-HT 的反向下调。
用不同浓度(0.05%至 0.2%v/v)的酒精处理树突状细胞 24-72 小时,并用 Q-PCR 和 Western blot 分析处理 SERT 和 MAO-A 的表达。此外,还使用 4-[4-(二甲氨基)-苯乙烯基]-1-甲基吡啶鎓碘化物摄取测定法测量在酒精处理的 DC 中 SERT 的功能,存在和不存在 SERT 抑制剂丙咪嗪。还使用 ELISA 定量测定培养上清液和细胞内 5-羟吲哚乙酸(5-HIAA)和环磷酸腺苷(cAMP)中的 5-HT 水平。
与未经处理的 DC 相比,用 0.1%酒精处理 24 小时的树突状细胞显示 SERT 和 MAO-A 的表达显著上调。我们还观察到,与未经处理的 DC 培养物相比,0.1%酒精增强了 SERT 的功能并降低了细胞外 5-HT 水平,这与细胞内 5-HIAA 和 cAMP 水平的升高有关。
我们的研究表明,酒精通过升高环磷酸腺苷来上调 SERT 和 MAO-A,这可能导致细胞外介质中 5-HT 浓度降低。由于 5-HT 是主要的神经递质和炎症介质,其酒精介导的消耗可能导致神经和免疫调节紊乱。