Proinflammatory effects on monocytes and Kupffer Cells in patients with Nonalcoholic Fatty Liver Disease (NAFLD)

INZAUGARAT ME; BALLESTA M; BAZ P; FONTANA D; LINARES M; SCIANGULA M; BECERRA GUTIERREZ C; GONZALEZ BALLERGA E; DARUICH J; SORDA JA; CHERÑAVSKY AC

SAN DIEGO, CA

Congreso; Digestive Disease Week; 2012

AASLD

Background and aims: Non alcoholic fatty liver diseases (NAFLD) include obesity-related disorders characterized by dislipidemia and hyperleptinemia. Reactive oxygen species (ROS) and cytokines production by Kuppfer Cells (KC) and monocytes (Mo) associate with inflammation and liver damage. We aimed to evaluate the effects of linoleic acid (LA) and leptin (Lep) on ROS and tumor necrosis factor (TNF)-A production in Mo and KC and to evaluate the potential anti-inflammatory effect of the antioxidant curcumin (CUR). Patients and methods: peripheral blood mononuclear cells (PBMC) and liver biopsies were obtained from adults with NAFLD (NAFLD, n=11) or individuals with liver diseases non NAFLD (non-NAFLD; n=6). PBMC from 30 healthy controls (Co) were also included. To evaluate ROS production, PMBC or liver cell suspensions obtained by chemical and mechanical methods were incubated with dichlorofluorescein-diacetate [5 µM], stimulated with Lep[10nM] or LA [200µM] +/-Cur[30µM], stained with anti-CD14 and -CD11b mAbs and analyzed by Flow Cytometry. A stimulation index (SI) results from the mean fluorescence intensity (MFI) in stimulated / unstimulated cells. Intracellular TNF-A was evaluated in Lep- stimulated [CD14+ TNF-A]PBMC and a fold of increase index (FI) was calculated. Kruskal-Wallis, Mann-Whitney with Bonferroni´s correction (pc), Wilcoxon matched pair test (pp) and Spearman´s rank correlation were conveniently used. Results: LA induced a higher production of ROS in Mo from NAFLD (pc=0.032 vs. Co) which diminished in the presence of CUR (pp= 0.030). LA similarly stimulated the oxidative burst of KC from patients with NAFLD (pp=0.015) and non-NAFLD (pp=0.04). Interestingly, ROS production induced by LA in both KC and Mo tend to correlate within NAFLD (r=0.75, p=0.06). Leptin induced a high production of TNF-A by Mo in Co, NAFLD and non-NAFLD (pp=0.031; 0.015 and 0.043 respectively) with a higher FI in NALFD (pc=0.029 vs. Co). Though the SI in Lep-stimulated Mo did not differ between groups, NAFLD patients show a tendency to a positive correlation between ROS and TNF-A production. Conclusions: A proinflammatory effect of LA and Lep on hepatic and peripheral myeloid cells is involved in NAFLD. A blockade of the proinflammatory response with curcumin might contribute to NAFLD treatment.