Materials and methods Animal treatments and sampling. Male mice (ddY, 3-week-old) were purchased from Seac Yoshitomi, Ltd. (Fukuoka, Japan) and were fed a commercial diet (Clea, Tokyo, Japan) and tap water ad libitum. They were divided into six groups from A to F, as shown in Fig. 1. The mice in groups C, D, E, and F were free to drink a 12% ethanol solution instead of tap water during the
apexbio indicated in Fig. 1. The mice in all groups were sacrificed at 10 months after the beginning of the experiment, and the livers were removed and analyzed for the experiment. Fig. 1. The experimental protocol. (A) Control [ND/water] for 10 months; (B) [ND/water] for the
decay series first 2 months and [0.06% 3′-MeDAB/water] for the last 8 months; (C) [ND/alcohol] for 10 months; (D) [ND/alcohol] for the first 2 months and [0.06% 3′-MeDAB/alcohol] for the last 8 months; (E) [ND/alcohol] for the first 2 months, [0.06% 3′-MeDAB/alcohol] for the next 4 months, and [0.06% 3′-MeDAB/water] for the last 4 months; (F) [ND/water] for the first 2 months, [3′-MeDAB/water] for the next 4 months, and [3′-MeDAB/alcohol] for the last 4 months. ND, normal diet; DAB, 3′-MeDAB.
