Hamby-Mason R, Chen J J, Schenker S, Perez A, Henderson G I
Department of Medicine, University of Texas Health Science Center at San Antonio 78284-7878, USA.
Alcohol Clin Exp Res. 1997 Sep;21(6):1063-72.
Fetal ethanol (E) exposure has well documented deleterious effects on brain development, yet it is uncertain if the neurotoxicity of maternal E consumption is generated by E itself, by its primary metabolite acetaldehyde (AcHO), or both. The current studies present evidence that homogenates of immature rat brains can generate AcHO via a catalase (CAT)-mediated reaction and that AcHO may be produced in vivo by this system. Homogenates of day 19 fetal rat brain were incubated with E (50 mM). When incubated with CAT inhibitors (sodium azide or 3-aminotriazole), AcHO formation was blocked, whereas neither the alcohol dehydrogenase inhibitor, 4-methylpyrazole, nor P-450 inhibitors decreased AcHO production. Three hours after one oral dose of E (4 g/kg) to a pregnant dam (gestation day 19), AcHO levels in fetal brain increased to 14.28 +/- 1.82 nM/g tissue. Baseline CAT activity in day 19 fetal brains was 4.5 times adult values (p < 0.05). Western blot analysis determined that CAT protein level in the day 19 fetal brain exceeded that in adult brain by 2.5 times. One hour after a single dose of E, CAT activity in day 19 fetal brain increased by 8.2 units/mg protein. In 5-day-old neonatal brains during the "third trimester" brain growth spurt, baseline CAT activity was twice the adult values (p < 0.05) and a 2-day in vivo E regimen increased AcHO levels to four times the control values, with a concomitant 1.7-fold increase in CAT activity. This was prevented by administration of a CAT inhibitor (3-amino-1,2,4-triazole). Immunohistochemical staining of neonatal brains exposed to E illustrated the presence of acetaldehyde-protein adducts. We conclude that AcHO is likely produced in rat fetal and neonatal brain via CAT-mediated oxidation of E. This phenomenon may be an important factor in the neurotoxic effects of in utero E exposure.
胎儿酒精(E)暴露对大脑发育具有已被充分记录的有害影响,但尚不确定母体摄入E后的神经毒性是由E本身、其主要代谢产物乙醛(AcHO)还是两者共同产生的。当前研究提供的证据表明,未成熟大鼠脑匀浆可通过过氧化氢酶(CAT)介导的反应生成AcHO,且该系统可能在体内产生AcHO。将第19天胎鼠脑匀浆与E(50 mM)一起孵育。当与CAT抑制剂(叠氮化钠或3-氨基三唑)一起孵育时,AcHO的形成被阻断,而酒精脱氢酶抑制剂4-甲基吡唑和P-450抑制剂均未降低AcHO的产生。在妊娠第19天给怀孕母鼠口服一剂E(4 g/kg)后3小时,胎脑中的AcHO水平升至14.28±1.82 nM/g组织。第19天胎脑的基线CAT活性是成年值的4.5倍(p<0.05)。蛋白质免疫印迹分析确定,第19天胎脑中的CAT蛋白水平比成年脑高出2.5倍。单次给予E后1小时,第19天胎脑中的CAT活性增加了8.2单位/mg蛋白。在“孕晚期”脑生长突增期间的5日龄新生鼠脑中,基线CAT活性是成年值的两倍(p<0.05),为期2天的体内E给药方案使AcHO水平增至对照值的四倍,同时CAT活性增加了1.7倍。给予CAT抑制剂(3-氨基-1,2,4-三唑)可防止这种情况发生。对暴露于E的新生鼠脑进行免疫组织化学染色显示存在乙醛-蛋白质加合物。我们得出结论,AcHO可能通过CAT介导的E氧化在大鼠胎儿和新生鼠脑中产生。这种现象可能是子宫内E暴露产生神经毒性作用的一个重要因素。