Carter E A, Wands J R
Department of Medicine, Harvard Medical School, Boston, Massachusetts.
Alcohol Clin Exp Res. 1988 Aug;12(4):555-62. doi: 10.1111/j.1530-0277.1988.tb00241.x.
We have studied a short term effect (96 hr) of ethanol on hormone-stimulated DNA synthesis in a primary rat hepatocyte culture system. Studies were also performed with respect to total RNA and protein synthesis as well as albumin secretion measured by a solid-phase radioimmunoassay. We found that ethanol, when added to cultured hepatocytes, resulted in a substantial reduction in hormone-stimulated hepatocyte DNA synthesis and this effect was concentration dependent and occurred in serum-free medium. Ethanol also had an inhibitory effect on total RNA synthesis but protein synthesis and albumin secretion remained essentially unchanged. We determined that hepatocytes exposed to ethanol during the first 24 hr of culture were the most susceptible to inhibition of DNA synthesis. During the first 24 hr, alcohol dehydrogenase (ADH) activity was present in the cells at higher levels than at 48 and 72 hr. In human hepatoma cell lines and differentiated primary and secondary chick fibroblasts, no ADH activity was demonstrable; such cells were not inhibited by 100 mM ethanol additions and DNA synthesis rates were similar to untreated cultures. Other alcohols found to be metabolized by hepatocyte ADH were inhibitory towards hormone-stimulated DNA synthesis whereas those with less metabolism had little effect. Hepatocytes treated with 4-methylpyrazole, an inhibitor of ADH, were partially protected from ethanol effects. Taken together our results are consistent with the hypothesis that a major physiological effect of ethanol on the hepatocyte is a direct impairment of DNA synthesis and that alcohol metabolism is required.
我们研究了乙醇对原代大鼠肝细胞培养系统中激素刺激的DNA合成的短期影响(96小时)。还针对总RNA和蛋白质合成以及通过固相放射免疫测定法测量的白蛋白分泌进行了研究。我们发现,当将乙醇添加到培养的肝细胞中时,激素刺激的肝细胞DNA合成会大幅减少,这种效应具有浓度依赖性,且在无血清培养基中也会出现。乙醇对总RNA合成也有抑制作用,但蛋白质合成和白蛋白分泌基本保持不变。我们确定,在培养的前24小时暴露于乙醇的肝细胞对DNA合成抑制最为敏感。在最初的24小时内,细胞中的乙醇脱氢酶(ADH)活性水平高于48小时和72小时时。在人肝癌细胞系以及分化的原代和二代鸡成纤维细胞中,未检测到ADH活性;添加100 mM乙醇不会抑制此类细胞,其DNA合成速率与未处理的培养物相似。其他被发现可被肝细胞ADH代谢的醇类对激素刺激的DNA合成具有抑制作用,而代谢较少的醇类则影响较小。用ADH抑制剂4-甲基吡唑处理的肝细胞可部分免受乙醇的影响。综合来看,我们的结果与以下假设一致:乙醇对肝细胞的主要生理作用是直接损害DNA合成,且需要酒精代谢。