Cardiovascular disease is one of the major causes of illness and death in developed countries. Myocardial infarcts result from the atherosclerotic process by the initiation from the arterial 'response to injury' phenomenon. Specific sites of the vascular tree are predisposed to protective �neointima� and subsequently develop fibro-fatty intimal lesions. However, such protective response can also contribute to the narrowing of arteries and the formation of hazardous plaques that can break out and cause blockages in vascular vessels. Different factors including diets can affect or enhance the formation of atherosclerotic lesions. For instance, low carbohydrate high protein (LCHP) diet was recently shown to induce more aortic atherosclerotic lesions and impaired angiogenesis compared to Western-type high fat diets. In the present study, a global metabolomics and lipidomics approach was employed to scrutinize the mechanisms of the lesion formation in ApoE-/- mice treated with LCHP, western and control diets. The extraction of compounds from plasma and aorta tissue samples was performed by high resolution mass spectrometry on LTQ Orbitrap Velos. For lipidomics analysis of plasma and aorta tissue, the performance of three sample prepration methods was systematically evaluated: (i) methanol/chloroform extraction, (ii) methyl tert-butyl ether liquid-liquid extraction and (iii) solid-phase microextraction (SPME). The methods were compared in terms of metabolite coverage, reproducibility, recovery of major lipid classes and ionization suppression. The ability of SPME to provide additional information (free concentration) was also investigated. All samples were analysed using three LC-MS methods (reversed-phase pentafluorophenyl, hydrophilic interaction liquid chromatography on underivatized silica and charged surface C18) in order to define and identify the most up- and down-regulated metabolites for each diet. Plasma profiles were finally correlated to the plaque composition observed for the three different diets.

Comparison of sample preparation methods for LC-MS metabolomics and lipidomics profiles of aortic tissue in apolipoprotein E-knockout mice: Combining analytical tools to examine the effect of low carbohydrate high protein diet

248th National Meeting (2014)

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