Fujimiya Tatsuya, Shinagawa Hiromi, Ohbora Yumiko, Aki Toshihiko
Department of Legal Medicine, Yamaguchi University School of Medicine, Minami-kogushi 1-1-1, Ube, Yamaguchi 755-8505, Japan.
Leg Med (Tokyo). 2003 Mar;5 Suppl 1:S123-5. doi: 10.1016/s1344-6223(02)00081-0.
We examined the disposition of ethanol, acetaldehyde and acetate in culture cells. Rat hepatoma-derived cells, H4IIE, were grown in T-shaped flasks to confluence in minimal essential medium supplemented with 5% fetal bovine serum. For the experimental group, the medium in the flask was replaced with a medium containing a 0, 4, 8 or 20 mM ethanol solution. For the control group, cell-free flasks containing media of the same ethanol concentrations were used. The ethanol, acetaldehyde and acetate concentrations in the flask were measured periodically by headspace gas chromatography. We found that the ethanol concentration of the medium decreased in both the control and the experimental groups; however, the rate of ethanol disappearance differed between these two groups, suggesting that ethanol is metabolized in the culture cells, and that its metabolism is saturated at a lower concentration. The acetaldehyde level was usually markedly low. Acetate was produced both with and without ethanol treatment, but the rate of acetate production increased with ethanol treatment, indicating that a certain percentage of acetate leaks into the medium. These findings suggest that ethanol in culture cells behaves in a kinetic fashion in vivo.
我们研究了培养细胞中乙醇、乙醛和乙酸盐的代谢情况。从大鼠肝癌中分离得到的细胞H4IIE，在补充有5%胎牛血清的最低必需培养基中于T形烧瓶中培养至汇合。对于实验组，将烧瓶中的培养基替换为含有0、4、8或20 mM乙醇溶液的培养基。对于对照组，使用含有相同乙醇浓度培养基的无细胞烧瓶。通过顶空气相色谱法定期测量烧瓶中乙醇、乙醛和乙酸盐的浓度。我们发现，对照组和实验组中培养基的乙醇浓度均下降；然而，这两组之间乙醇消失的速率不同，表明乙醇在培养细胞中被代谢，并且其代谢在较低浓度下达到饱和。乙醛水平通常明显较低。无论有无乙醇处理都会产生乙酸盐，但乙醇处理会使乙酸盐的产生速率增加，表明一定比例的乙酸盐泄漏到培养基中。这些发现表明，培养细胞中的乙醇在体内表现出动力学行为。
