Resveratrol res

ZTC and resveratrol were dissolved in water for intragastric administration at a volume of 0.1 ml per 10 g body weight. Different doses of ZTC were given continuously for 8 days via intragastric administration, and 5 mg/kg of LPS was administered intraperitoneally 1 h after the last day of administration. The mice were grouped based on the administration of ZTC dosages as follows: Normal group, 0.9% normal saline; model group (LPS), 5 mg/kg (Qin et al., 2013 (link)); positive group (resveratrol RES, Sigma-Aldrich, St. Louis, MO), 40 mg/kg, and the detailed administration of RES is described as: mice receive daily intraperitoneal injections of RES that is dissolved in 0.2 ml of 2% ethyl alcohol (Maheedhar et al., 2015 (link)); ZTC low dose group, (L) 0.17 g/kg; ZTC medium dose group, (M) 0.34 g/kg; ZTC high dose group, (H) 0.7 g/kg).

Alpha mouse liver 12 (AML12) cells were purchased from Procell Life (CL-0602, Wuhan, China). The AML12 cells were cultured in DMEM/F12 (with 10% FBS, 1% penicillin/streptomycin, 10 μg/mL insulin, 5.5 μg/mL transferrin, 5 ng/mL selenium, and 40 ng/mL dexamethasone) in a humidified environment at 37 °C and 5% CO₂. Unconjugated bilirubin (UCB) (14370, Sigma-Aldrich, St. Louis, MO, USA) was utilized in the treatment of cells, and was prepared according to the article by Granato et al.^{57 (link)}. The UCB solution was diluted with DMEM/F12 medium containing 5% FBS to prepare working solutions of 50 μM, 100 μM, and 200 μM. The molar ratios of UCB to albumin were 2.75, 5.5, and 11, respectively. In this study, OA, BMS-986020 (BMS) (HY-100619, Med Chem Express, NJ, USA), MK-571 and resveratrol (Res) (R5010, Sigma-Aldrich, St. Louis, MO, USA) were added 2 h before UCB (100 μM) administration.

Mouse monoclonal antibodies against FLAG (Sigma, F1804), Myc (CST, 2276S), HCV NS3 (Abcam, ab65407) and Core (Thermo, MA1-080), rabbit monoclonal antibody against HA (CST, 3724S), QPRT (Abcam, ab180930), Smurf2 (CST, 12024), rabbit polyclonal antibody against SREBP-2 (Santa Cruz, sc-13552), and fine chemicals of proteasome inhibitor MG132 (ApexBio Tech, A2585), 2,3-Pyridinedicarboxylic acid (QA, Sigma, P63204), 2,3-Pyridinedicarboxylic Acid-d3 (QA-d3, J&K Scientific Ltd, P991633), β-Nicotinamide adenine dinucleotide hydrate (NAD, Sigma, N7004), Resveratrol (Res, Sigma, V900386), Phthalic acid (PHT, Sigma, 80010), Clofibrate (Sigma, C6643) and cycloheximide (Sigma, C7698) were purchased from where indicated.

Human TCC EJ cells [13] (link) were cultured in Dulbecco’s modified Eagle’s essential medium (DMEM) containing 10% fetal bovine serum (Gibco Life Science, Grand Island, NY, USA) under 37°C and 5% CO₂ conditions. The cells (5×10⁴/ml) were plated to culture dishes (NUNC, Denmark) and incubated for 24 h before the experiments.
Resveratrol (Res; Sigma Chemical, Inc, St. Louis, MO) was dissolved in dimethylsulfoxide (DMSO; Sigma) and diluted with culture medium to the working concentrations just before use. The cells under normal culture condition, treated by 0.2% DMSO and exposed to 100 µM Res for 48 h were used as normal, background and efficacy controls, respectively. As shown in the diagram (Figure 1A), EJ cells were treated by 100 µM, 150 µM or 200 µM Res for 1 h, 1.5 h or 2 h in 24 h intervals. After 1 h and 2 h treatments, Res containing media were replaced with normal medium upon 3 washes. Therefore, EJ cells were exposed to different concentrations of Res for 3 times (once a day) during the 72 h experiment (Figure 1A). Cell numbers and viabilities were checked in 12 h intervals. The cell-bearing coverslips were fixed in cold acetone or 4% paraformaldehyde (pH 7.4) for morphological and immunocytochemical examinations. The experimental groups were set in triplicate and the experiments were repeated for three times to establish confidential conclusion.