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Nutrient Interactions and Toxicity

Heart 7-Hydroperoxycholesterol and Oxysterols Are Elevated in Chronically Ethanol-Fed Rats¹

Junko Adachi^2,*, Risa Kudo^*, Yasuhiro Ueno^*, Ross Hunter, Rajkumar Rajendram, Elizabeth Want and Victor R. Preedy

Department of Legal Medicine, Kobe University Graduate School of Medicine and Departments of Clinical Biochemistry and Nutrition and Dietetics, King’s College London, London, U.K. ^*

²To whom correspondence should be addressed. E-mail: adachi{at}med.kobe-u.ac.jp

ABSTRACT

Recently, cholesterol hydroperoxides have been shown to be sensitive pathogenic markers of reactive oxygen species (ROS)-mediated damage though they have never been measured in heart tissue. We hypothesized that cholesterol hydroperoxides and oxysterols, putative cardiotoxic products of cholesterol oxidation, are elevated in the hearts of alcoholics as a consequence of ROS-mediated reactions. To test this, we measured 7- and 7ß-hydroperoxycholest-5-en-3ß-ol (7-OOH and 7ß-OOH) by HPLC with postcolumn chemiluminescence as well as 7- and 7ß-hydroxycholesterol (7-OH and 7ß-OH) and 3ß-hydroxycholest-5-en-7-one (also termed 7-ketocholesterol; 7-keto) by HPLC-UV in cardiac muscle of alcohol-fed rats. Alcohol feeding was carried out using a pair-feeding protocol with 35% of total dietary energy as ethanol; controls were pair-fed isocaloric glucose. After 6–7 wk treatment with alcohol, heart 7-OOH, 7ß-OOH and 7ß-OH were significantly greater than in controls. Levels of heart phospholipid 16:0 and 18:1 were lower than in controls, while 18:0 and 18:2 were greater. This is the first report of the presence of 7-OOH, 7ß-OOH and 7-OH in cardiac tissue. The elevations in 7-OOH and 7ß-OOH as well as 7ß-OH are evidence of increased oxidative stress and possible membrane changes. Alterations in the proportions of 16:0, 18:1, 18:2 and 18:0 in heart phospholipids provide further evidence of an altered membrane domain.

KEY WORDS: • 7-hydroperoxycholesterols • oxysterols • ethanol • rats • heart

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