Pigs fed saturated fat/cholesterol have a blunted hypothalamic-pituitary-adrenal function, are insulin resistant and have decreased expression of IRS-1, PGC1α and PPARα.

The increasing incidence of insulin resistance has been linked to both increased intake of saturated fatty acids and disruption of the hypothalamic-pituitary-adrenal (HPA) axis. We tested the hypothesis that adding saturated fat/cholesterol to the diet of growing pigs would both disrupt HPA function and cause insulin resistance. Three-month-old pigs were fed either a control (13% energy from fat) or a high saturated fatty acid cholesterol (HSFC) diet (44% energy from fat; 2% cholesterol). After 10 weeks on the diets, intravenous ACTH, insulin and glucose challenges were performed, and after 12 weeks, tissue samples were taken for measurement of mRNA and for lipid-rich aortic lesions. Plasma total, HDL- and LDL-cholesterol were significantly increased in pigs fed the HSFC diet. Cortisol release during the ACTH challenge was suppressed in HSFC-fed pigs which were also more insulin resistant and glucose intolerant than controls. The HSFC diet decreased the expression of insulin receptor (IR) and insulin receptor substrate-1 in muscle and adipose tissue as well as adiponectin and adiponectin receptor 2 expression in fat. The HSFC diet decreased PGC-1α and PPARα expression in muscle but increased PPARα expression in liver. There was a trend for an increase in lipid-stained lesion frequency around the abdominal branches of the aorta in HSFC-fed pigs. We conclude that feeding increased saturated fat to pigs causes disruption in the HPA axis, insulin resistance and decreased muscle and adipose expression of genes controlling insulin signalling and mitochondrial oxidative capacity.

Effect of aortic taper on patterns of blood flow and wall shear stress in rabbits: association with age.

OBJECTIVE: The distribution of atherosclerotic lesions changes with age in human and rabbit aortas. We investigated if this can be explained by changes in patterns of blood flow and wall shear stress. METHODS: The luminal geometry of thoracic aortas from immature and mature rabbits was obtained by micro-CT of vascular corrosion casts. Blood flow was computed and average maps of wall shear stress were derived. RESULTS: The branch anatomy of the aortic arch varied widely between animals. Wall shear was increased downstream and to a lesser extent upstream of intercostal branch ostia, and a stripe of high shear was located on the dorsal descending aortic wall. The stripe was associated with two vortices generated by aortic arch curvature; their persistence into the descending aorta depended on aortic taper and was more pronounced in mature geometries. These results were not sensitive to the modelling assumptions. CONCLUSIONS: Blood flow characteristics in the rabbit aorta were affected by the degree of taper, which tends to increase with age in the aortic arch and strengthens secondary flows into the descending aorta. Previously-observed lesion distributions correlated better with high than low shear, and age-related changes around branch ostia were not explained by the flow patterns.

Atheroprotective effects of dietary L-arginine increase with age in cholesterol-fed rabbits.

NO has several putative atheroprotective properties but its precursor, L-arginine, and inhibitors of its synthesis have had inconsistent effects on the extent of experimental atherosclerosis in rabbits. The location and character of experimental atherosclerosis differ between immature and mature rabbits; both phenomena have been attributed to changes with age in the NO pathway. We investigated whether the influence of dietary L-arginine on experimental atherosclerosis is also age-related. The frequency of lesions was mapped in the descending thoracic and upper abdominal aorta of immature and mature rabbits fed 1 % cholesterol, with or without supplementary L-arginine, for 8 weeks. Consistent with earlier data, the distribution of lesions around the branch points changed with age in control rabbits. The mean frequency of lesions was essentially the same at both ages. L-Arginine supplements had no effect on the distribution of lesions at either age. They significantly reduced the mean frequency of lesions in mature animals but not in immature animals. Thus, the atheroprotective effect of dietary L-arginine in cholesterol-fed rabbits increases with age.

Application of Fourier transform infrared spectroscopic imaging to the study of effects of age and dietary L-arginine on aortic lesion composition in cholesterol-fed rabbits.

Diet-induced atherosclerotic lesions in the descending thoracic segment of rabbit aorta were analysed ex vivo by micro-attenuated total reflection (ATR)-Fourier transform infrared (FTIR) spectroscopic imaging. The distribution and chemical character of lipid deposits within the arterial wall near intercostal branch ostia were assessed in histological sections from immature and mature rabbits fed cholesterol with or without l-arginine supplements. Previous studies have shown that both these properties change with age in cholesterol-fed rabbits, putatively owing to changes in the synthesis of nitric oxide (NO) from l-arginine. Immature animals developed lesions at the downstream margin of the branch ostium, whereas lipid deposition was observed at the lateral margins in mature animals. Dietary l-arginine supplements had beneficial effects in mature rabbit aorta, with overall disappearance of the plaques; on the other hand, they caused only a slight decrease of the lipid load in lesions at the downstream margin of the ostium in immature rabbits. ATR-FTIR imaging enabled differences in the lipid to protein density ratio of atherosclerotic lesions caused by age and diet to be visualized. Lipid deposits in immature rabbits showed higher relative absorbance values of their characteristic spectral bands compared with those in immature l-arginine-fed rabbits and mature rabbits. The multivariate methods of principal component analysis (PCA) and factor analysis (FA) were employed, and relevant chemical and structural information were obtained. Two distinct protein constituents of the intima-media layer at different locations of the wall were identified using the method of FA. This approach provides a valuable means of investigating the structure and chemistry of complex heterogeneous systems. It has potential for in vivo diagnosis of pathology.

Use of a desktop scanner and spreadsheet software for mapping arterial disease.

Mapping the surface area and distribution of atherosclerosis has so far required photographic, video, or photomicrographic imaging of the inner surface of the arterial wall, as well as laborious manual or sophisticated image processing methods of quantification. We investigated whether comparable results could be obtained by using a conventional flatbed scanner and readily available spreadsheet software. Lipid-rich lesions near 21 aortic branches from cholesterol-fed rabbits were mapped using a scanner-based technique and an established photomicrographic technique. When the tissue was counterstained and held on the scanner by a transparent weight to obtain adequate contrast and prevent detection of adventitial staining, the areas of lipid deposition detected by the two methods correlated highly (R2 = 0.99). Discrepancies arose mainly at the edges of lesions, probably because of alignment errors and better flattening of the tissue on the scanner; when these were accounted for, discrepancies occurred in < 1% of the total area examined. The new method produces results comparable with previous procedures, but is much more rapid and requires only office equipment and software.

Evidence for a reversal with age in the pattern of near-wall blood flow around aortic branches.

The changes that occur with age in the distribution of atherosclerotic lesions around arterial branch points challenge accepted theories relating disease to haemodynamic stresses. We investigated whether flow near branch points changes with age in a way that can account for the different lesion distributions. Flow around 20 branches from immature and mature aortas was investigated by examining the length:width ratio and orientation of endothelial nuclei; these properties depend on the magnitude and direction of near-wall flows, respectively. There were significant changes in the pattern of nuclear shape with age, consistent with a reversal in the pattern of shear around branches. In control regions away from branches, there were no such changes. The role of haemodynamic stresses in atherogenesis may require re-evaluation in the light of these results.