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This record contains raw data related to the article  “12(S)-Hydroxyeicosatetraenoic acid downregulates monocyte-derived macrophage efferocytosis: new insights in atherosclerosis"

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Abstract:The involvement of 12(S)-hydroxyeicosatetraenoic acid (12(S)-HETE), a 12-lipooxygenase product of arachidonic acid, has been suggested in atherosclerosis. However,its effecton macrophage functions is not completely understood, so far. The uptake of apoptotic cells (efferocytosis) by macrophages is an anti-inflammatory process, impaired in advanced atherosclerotic lesions. This process induces the release of the anti-inflammatory cytokine interleukin-10 (IL-10), and it is regulated by Rho-GTPases, whose activation involves the isoprenylation, a modification inhibited by statins. We assessed 12-HETE levels in serum of coronary artery disease (CAD) patients, and explored12(S)-HETE invitro effect on monocyte-derived macrophage (MDM) efferocytosis.Sixty-four CAD patients and 24 healthy subjects (HS) were enrolled. Serum12-HETE levels were measured using a tandem mass spectrometry method. MDMs, obtained from a spontaneous differentiation of adherent monocytes, were treated with12(S)-HETE (10–50  ng/mL). Efferocytosis and RhoA activation were evaluated by flow cytometry. IL-10 was measured by ELISA. CAD patients showed increased 12-HETE serum levels compared to HS (665.2 [438.1–896.2] ng/mL and 525.1 [380.1–750.1] ng/mL, respectively, p < 0.05) and reduced levels of IL-10. MDMs expressed the12(S)-HETE cognate receptor GPR31. CAD-derived MDMs displayed defective efferocytosis vs HS-MDMs (9.4 [7.7–11.3]% and 11.1 [9.6–14.1]% of MDMs that have engulfed apoptotic cells, respectively, p < 0.01). This reduction is marked in MDMs obtained from patients not treated with statin (9.3 [7.4–10.6]% statin-free CAD vs HS, p=0.01; and 9.9 [8.6–11.6]% statin-treated CAD vs HS, p=0.07). The in vitro treatment of MDMs with12 (S)-HETE (20ng/mL) induced 20% decrease of efferocytosis (p < 0.01) and 71% increase of RhoA activated form(p < 0.05). Atorvastatin (0.1μM) counteracted these 12(S)-HETE-mediated effects.These results show a12(S)-HETE pro-inflammatory effect and suggest a new potential contribution of this mediator in the development of atherosclerosis.

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