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Supplementation of Diets with the Black Rice Pigment Fraction Attenuates Atherosclerotic Plaque Formation in Apolipoprotein E Deficient Mice¹

Min Xia, Wen Hua Ling², Jing Ma, David D. Kitts^* and Jerzy Zawistowski^*,

Department of Clinical Nutrition, School of Public Health, Sun Yet-sen University (Northern Campus), Guangzhou, 510080, People’s Republic of China; ^* Food Science, Food Nutrition and Health Program, University of British Columbia, Vancouver, British Columbia, Canada V6T-1Z4 and Forbes Medi-Tech, Inc., Vancouver, British Columbia, Canada V6C-2T8

²To whom correspondence should be addressed. E-mail: whling{at}gzsums.edu.cn

Apolipoprotein (apo)E-deficient mice were used to study the antiatherogenic effect of black rice pigment fraction (BRF) and the possible mechanisms by which it inhibits atherogenesis. The apoE-deficient mice (n = 45) were randomly divided into three groups and received AIN-93G diet (positive group), AIN-93G with 5 g of black rice pigment fraction/100 g (BRF group) and AIN-93G with 5 g of white rice outer layer fraction/100 g (WRF group) for 16 wk. C57BL/6J mice (n = 15) received AIN-93G and were used as a control group. Blood samples were collected for measurement of lipid concentration, antioxidized LDL antibody and nitric oxide concentration. Livers were extracted for determination of cholesterol concentrations, and aortas were used to determine cholesterol concentrations and inducible nitric oxide synthase protein and mRNA expression. Hearts were used to assess atherosclerotic plaque formation. The apoE-deficient mice fed the black rice pigment fraction diet had 48% (P < 0.01) less atherosclerotic lesion area compared with apoE-deficient mice fed only the AIN-93G diet and 46% (P < 0.01) less lesion area compared with mice fed the white rice outer layer fraction diet. This observation corresponded with significantly (P < 0.05) lower total serum cholesterol, lower liver and aorta cholesterol (P < 0.01) and higher HDL cholesterol (P < 0.05) concentrations and lower (P < 0.05) antioxidized LDL antibody titer in apoE-deficient mice fed the black rice pigment fraction diet compared with positive and WRF groups. Notwithstanding this, mice fed the black rice pigment fraction diet also had lower CD4⁺ T lymphocyte expression (P < 0.05) and weaker inducible nitric oxide synthase expression (P < 0.05) compared with mice fed the AIN-93G diet and the white rice outer layer fraction diet, respectively. We concluded that the inhibition of atherosclerotic lesions of the black rice pigment fraction is attributed to the improvement in cholesterol accumulation and reduction in oxidative stress and inflammation.

KEY WORDS: • black rice pigment fraction • apoE-deficient mice • LDL oxidation • atherosclerosis • inflammatory response

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