Table 1. Mouse model for EtOH-induced liver disease and testing therapeutic agents, such as probiotics.
| Micea | Parab |
Mouse group |
Disease indicators | Materials and methods | |||||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| Conc | 4EtOHd | 8EtOH | 12EtOH | 4EtOH-P | 8EtOH-P | 12EtOH-P | 8EtOH-4P | ||||
| BALB/cAnNHsd; female; 12 months age; light–dark cycle 12–12 h; 23 °C; 4RF21 standard diet (Mucedola, Italy) | Food and water weight; body weight; caloric intake | +15% EtOH 4 weeks | +15% EtOH 8 weeks | +15% EtOH 12 weeks | +15% EtOH 4 weeks | +15% EtOH 8 weeks | +15% EtOH 12 weeks | +15% EtOH 8 weeks | Liver steatosis, fibrosis; iNOS levels, Hsp60 levels and gene expression, Hsp60 nitration | Liver tissue; histopathology, immunohistochemistry, qRT-PCR, western blotting, immunoprecipitation, statistical analysis | |
| +L.f. e 109 CFU | +L.f. 109 CFU | +L.f. 109 CFU | +L.f. 109 CFU 4 weeks | ||||||||
Abbreviations: CFU, colony-forming units; EtOH, ethanol; Hsp, heat-shock protein; iNOS, inducible form of nitric oxide synthase; L.f., Lactobacillus fermentum; qRT-PCR, real-time quantitative PCR.
Mice characteristics, environmental conditions, and basic diet.
Parameters measured initially and periodically thereafter (see text).
Control group receiving the standard diet only (no ethanol or probiotic).
For the experimental mice, EtOH was added to account for 15% of total calories in the diet. To acclimate the mice to the alcohol-containing diet, they were given 5% EtOH-calorie content for 2 days, then 10% EtOH-calorie content for 2 days, and lastly 15% EtOH-calorie content for 4, 8, and 12 weeks (mouse groups 4EtOH, 8EtOH, and 12EtOH, respectively). In addition, groups 4EtOH-P, 8EtOH-P, and 12EtOH-P received orally the probiotic L.f., 109 CFU per animal per day, 5 days per week until the end of the experiments. Group 8EtOH-4P received the ethanol-containing diet for 8 weeks as group 8EtOH, and then L.f., 109 CFU per animal per day, 5 days per week for 4 weeks. The pertinent EtOH dose was diluted in water to a final volume of 100 μl. This volume was orally administered to the mice every day using a pipette and a tip and, after 20 min, the mice were fed with the probiotic in the same manner.
L.f., a probiotic used to test the suitability of the model to detect effects, beneficial or otherwise, on the disease indicators. These groups can be used as positive controls and as the baseline for comparing efficacy when testing other probiotics or drugs.