We also report evidence of the induction of profibrotic pathways in association with histological evidence for hepatic fibrogenesis and raised NAFLD Activity Score (NAS). Mechanistic studies implicate key components of the hepatic innate immune system because NKT cell numbers were attenuated, whereas KCs, although increased in number, with evidence of enhanced ROS production, paradoxically had defective phagocytosis activity, as previously reported on in human NAFLD.18 Female C57BL/6J mice (proven breeders with one previous pregnancy,

n = 20; Charles River UK Ltd., Margate, UK) were fed standard chow (RM1) or a highly palatable obesogenic diet consisting of a semisynthetic energy-rich, high-fat diet (10% simple sugars, 18% animal lard, 4% soya oil, 28% polysaccharide, 23% protein [w/w]; diet code: 824053; 45% AFE FAT energy, 4.5 buy Dabrafenib kcal/g; n = 30; Special Dietary Services, Essex, UK), supplemented with fortified sweetened condensed milk (Nestle, SZ) for 6 weeks ad libitum, as previously described.3 Final dietary composition based on intake was approximately 16% fat, 33% simple sugars, 15% protein, and energy

4.0 kcal/g. Mice on the obesogenic diet entered the breeding protocol after achieving a 30% increase in body weight, and controls were aged matched. All animals were treated in accord with the Animals (Scientific Procedures) Act (UK) 1986 guidelines. Pregnant dams were maintained on their respective Kinase Inhibitor Library purchase diets throughout pregnancy and lactation. After spontaneous delivery, litter sizes were standardized to 6 pups per litter. Female offspring born to

either lean or obese dams were weaned onto a standard chow diet (n = 10) (OffCon-SC or OffOb-SC) or the high-fat diet (diet code: 824053; 45% AFE FAT; n = 10; Special Dietary Services) (OffCon-OD or OffOb-OD). After sacrifice, offspring body weight, inguinal fat pad mass, and hepatic tissue TG content were determined at 3 months of age. Markers of liver injury, fibrogenesis, and liver histology were assessed at 3 and 12 months. Investigation of the hepatic innate immune system was completed at 6 months. Blood was 上海皓元 collected by cardiac puncture and plasma was assayed for ALT by the Royal Free Hospital Clinical Biochemistry Department (London, UK). Whole liver tissue TG was determined by an adaptation of the Folch Method19 and an enzymatic colorimetric assay (UNIMATE 5 TRIG, Roche BC1; Roche Diagnostics, Sussex, UK). Real-time polymerase chain reaction (PCR) was performed using the QuantiTect SYBR Green PCR System with HotStar Taq DNA Polymerase (Qiagen, Hilden, Germany). Gene-specific primers were designed for IL-6, IL-12, IL-18, TNF-α, alpha smooth muscle actin (α-SMA), transforming growth factor-β (TGF-β), and collagen type 1 alpha 2 (Col1-α2), as previously reported.