Individually housed and body weight-matched 7-week-old C57BL/6J males (Harlan, the Netherlands) were treated with vehicle (amorphous nanoparticles with 3.4% dimethylacetamide), AZ876 (20 ��mol?kg?1?day?1, selleck chemicals llc ~9?mg?kg?1?day?1), or GW3965 (34 ��mol?kg?1?day?1, ~20 mg?kg?1?day?1) by oral gavage twice daily for 8 days. On day 6, labelled J774A1 cells (2.5 �� 106 viable cells containing 28 �� 106 dpm in 0.4 mL medium) were injected intraperitoneally. Faeces were collected between 24 and 48 h after macrophage injection. At termination (48 h after macrophage injection), the animals were anaesthetized with isoflurane (Forene, Abbot Scandinavia AB, Solna, Sweden), plasma was collected and the liver was collected after perfusion with PBS.

Faeces (total) and liver (100 mg) were homogenized in PBS and total lipids were extracted from faeces and liver homogenates and plasma according to Folch et al. (1957). Free cholesterol and cholesteryl ester were separated using thin layer chromatography (hexane : diethylether : acetic acid 90:10:1). [3H] in total extracted lipids, free cholesterol and cholesteryl ester were counted in a liquid scintillator and presented as percent dpm injected into the animal. The lipoprotein particles were separated using size-exclusion high-performance liquid chromatography and the [3H] was found in the HDL fraction (data not shown). Study of treatments for atherosclerosis Female heterozygous APOE*3Leiden transgenic mice (about 13 weeks of age, bred by TNO), on a C57bl/6 background and characterized by elisa for human apoE (van Vlijmen et al., 1994), were used.

During a 3 week run-in period, all animals received a semi-synthetic Western-type diet containing 40.5% sucrose, 15% cacao butter, 0.50% (w/w) cholesterol and 0.33% v/w sunflower oil (necessary to dissolve the compounds). After matching into four groups, based on age, plasma cholesterol and triglyceride levels, the mice received Western-type diet either alone (control group) or supplemented with AZ876 in a low dose (5 ��mol?kg?1?day?1; ~2 mg?kg?1?day?1) or high dose (20 ��mol?kg?1?day?1; ~9 mg?kg?1?day?1) or with GW3965 (17 ��mol?kg?1?day?1; ~10 mg?kd?1?day?1) (Joseph et al., 2002). Initially, the high dose of AZ876 was 10 ��mol?kg?1?day?1; however, after 4 weeks of treatment, GW3965 induced hypertriglyceridaemia but AZ876 (10 ��mol?kg?1?day?1) did not.

Therefore, it was decided to increase the dose of AZ876 to 20 ��mol?kg?1?day?1. Both compounds were provided by AstraZeneca (M?lndal, Sweden). After 20 weeks, mice were killed and the hearts, aortic root, livers Batimastat and small intestine (duodenum) were isolated. Plasma lipid and lipoprotein analysis and inflammation markers After a 4 h fasting period from 9 am to 1 pm, blood was collected and EDTA plasma was obtained subsequently (Sarstedt, N��mbrecht, Germany) and lipoproteins were separated by fast protein liquid chromatography (FPLC; Westerterp et al., 2006).