Sumida Ken D, Cogger Alma A, Matveyenko Aleksey V
Department of Biological Sciences, Chapman University, Orange, CA 92866, USA.
Alcohol. 2007 Mar;41(2):67-75. doi: 10.1016/j.alcohol.2007.02.002. Epub 2007 Apr 26.
The impact of alcohol-induced suppression on hepatic gluconeogenesis (HGN) after chronic ethanol consumption between males and females is unknown. To determine the effects of chronic alcohol consumption (8 weeks) on HGN, the isolated hepatocyte technique was used on 24 h fasted male and female Wistar rats. Livers were initially perfused with collagenase and the hepatocytes were isolated. Aliquots of the cell suspension were placed in Krebs-Henseleit buffer and incubated for 30 min with lactate, [U -14C]lactate, and nine different concentrations of ethanol (EtOH). Dose-effect curves were generated for the determination of maximal and half-maximal alcohol-induced inhibition on HGN. There was no significant difference in HGN (lactate only and no EtOH) between males and females fed the control diet (88.5 +/- 5.1 nmol/mg protein/30 min). Similarly, the HGN (lactate only and no EtOH) in males fed the ethanol diet (ME) were not significantly different (82.8 +/- 3.5 nmol/mg protein/30 min) compared to controls. In contrast, the females chronically fed the ethanol diet (FE) had significantly (P < .05) lower HGN (67.8 +/- 4.6 nmol/mg protein/30 min) compared to both ME and controls. With alcohol in the incubation medium, the HGN significantly (P<.05) declined in all groups. While alcohol suppressed HGN to a larger (P < .05) extent in ME (45.8 +/- 3.7 nmol/mg protein/30 min) compared to controls (64.0 +/- 3.8 nmol/mg protein/30 min), the inhibition was even greater (P < .05) in FE (32.7 +/- 3.2 nmol/mg protein/30 min). The more pronounced effect of chronic alcohol consumption on HGN in the presence of ethanol in female hepatocytes was supported by the concomitant decreases (P < .05) in 14C-lactate incorporation into 14C-glucose, lactate uptake, and 14C-lactate uptake. The results suggest that chronic alcohol consumption elicits a greater reduction on HGN in the presence of ethanol in the hepatocytes of females compared to males.
长期乙醇摄入后，酒精诱导的抑制作用对雄性和雌性肝脏糖异生（HGN）的影响尚不清楚。为了确定长期酒精摄入（8周）对HGN的影响，对禁食24小时的雄性和雌性Wistar大鼠采用分离肝细胞技术。肝脏先用胶原酶灌注，然后分离肝细胞。将细胞悬液的等分试样置于Krebs-Henseleit缓冲液中，与乳酸、[U-14C]乳酸和九种不同浓度的乙醇（EtOH）一起孵育30分钟。生成剂量效应曲线，以确定酒精诱导的对HGN的最大抑制和半最大抑制。喂食对照饮食的雄性和雌性之间的HGN（仅乳酸，无EtOH）没有显著差异（88.5±5.1 nmol/mg蛋白质/30分钟）。同样，喂食乙醇饮食（ME）的雄性的HGN（仅乳酸，无EtOH）与对照组相比没有显著差异（82.8±3.5 nmol/mg蛋白质/30分钟）。相比之下，长期喂食乙醇饮食的雌性（FE）的HGN（67.8±4.6 nmol/mg蛋白质/30分钟）与ME组和对照组相比显著降低（P<.05）。在孵育培养基中加入酒精后，所有组的HGN均显著下降（P<.05）。与对照组（64.0±3.8 nmol/mg蛋白质/30分钟）相比，酒精对ME组HGN的抑制作用更大（P<.05）（45.8±3.7 nmol/mg蛋白质/30分钟），而在FE组中抑制作用甚至更大（P<.05）（32.7±3.2 nmol/mg蛋白质/30分钟）。14C-乳酸掺入14C-葡萄糖、乳酸摄取和14C-乳酸摄取的同时减少（P<.05），支持了长期酒精摄入对雌性肝细胞中存在乙醇时HGN的更显著影响。结果表明，与雄性相比，长期酒精摄入在雌性肝细胞中存在乙醇时对HGN的降低作用更大。
