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The present study was aimed to investigate the effect of 12 weeks of extra-virgin olive oil (EVOO) consumption on the capacity of HDL to promote cholesterol efflux (CE) and to determine which CE pathways are modulated by EVOO consumption. Whole HDL and HDL 2 /HDL 3 subclasses were isolated from the plasma of twenty-six healthy volunteers before and after 12 weeks of EVOO consumption (25 ml/d). EVOO consumption increased the capacity of serum and HDL to mediate CE from THP-1, J774 macrophages and Fu5AH cells by 9·8-24·57 %, depending on the cell type. The increase in CE was independent of both HDL concentration and subclass distribution. The three HDL-mediated CE pathways (ATP-binding cassette (ABC) A1, ABCG1 and scavenger receptor class B type I (SR-BI)) were modulated by EVOO consumption. The fluidity of the phospholipidic layer of HDL increased by 13 % (P,0·001) following EVOO consumption compared with baseline. EVOO consumption also increased the release of excess cholesterol from human monocyte-derived macrophages (HMDM) by 44 % (P, 0·001), and ABCA1 and ABCG1 mRNA transcription by 16·08 % (P, 0·001) and 35·79 % (P, 0·01), respectively. The protein expression of these two cholesterol transporters also increased after EVOO consumption. In contrast, SR-BI mRNA and protein expression in HMDM were significantly lower after 12 weeks of EVOO consumption. Incubating J774 macrophages with EVOO polyphenol extracts induced a concentration-dependent up-regulation of ABCA1 and ABCG1 expression in macrophages. After 12 weeks of EVOO consumption, the capacity of HDL to mediate CE was improved and the ability of HMDM to release excess cholesterol was enhanced by increasing the expression of ABCA1 and ABCG1 transporters.

Paraoxonase 1 (PON1) is associated with HDL and modulates the antioxidant and anti-inflammatory role of HDL. The goals of the present study were to investigate the effect of ageing and the role of PON1 on the anti-inflammatory activity of HDL, and to determine whether extra-virgin olive oil (EVOO) consumption could improve the atheroprotective activity of HDL. HDL and PON1 were isolated from the plasma of ten young (Y-HDL and Y-PON1) and ten elderly (E-HDL and E-PON1) healthy volunteers before and after 12 weeks of EVOO consumption. Inflammation was assessed by measuring intracellular adhesion molecule 1 (ICAM-1) expression. THP-1 (human acute monocytic leukaemia cell line) monocyte chemotaxis was measured using a Boyden chamber. Oxidative damage to HDL was assessed by measuring conjugated diene formation and changes in electrophoretic migration. Y-HDL had more anti-inflammatory activity than E-HDL. The conjugated diene content and the electrophoretic mobility of E-HDL were higher than those of Y-HDL. Y-PON1 had significant anti-inflammatory activity, reducing ICAM-1 expression by 32·64 (SD 2·63) %, while E-PON1 had no significant effect. THP-1 chemotaxis measurements confirmed the ICAM-1 expression results. The 12 weeks of EVOO consumption significantly increased the anti-inflammatory activities of both HDL and PON1. The anti-inflammatory activity of HDL was modulated by PON1 and was lower in the elderly volunteers. EVOO consumption increased the anti-inflammatory effect of HDL and reduced the age-related decrease in anti-atherogenic activity.

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