W1250 Effect of Nutritional Supplementation with Zinc and Vitamin C in Acute Hemorrhagic Rectal Ulcer Patients: A Randomized Clinical Trial

Abstract

Introduction: Green tea polyphenols, particularly -(-) epigallocatechin 3-gallate (EGCG), have sparked considerable interest as potential therapeutic agents for chronic inflammatory diseases due to their anti-oxidant and anti-inflammatory effects. Aim: Analyze the ability of EGCG to inhibit production of pro-inflammatory cytokines in human peripheral blood mononuclear cells (PBMCs) In Vitro to gauge its anti-inflammatory effects in immune cells from IBD patients. Methods: PBMCs were obtained from 4 colitis patients and 4 normal controls and enriched for monocytes or lymphocytes by plastic adherence. CD14+ macrophages, CD4+CD45+RO+ and CD4+CD45+RA+ T cells were isolated respectively by microbead technology. Three experiments were performed: 1) Lipopolysaccharide (LPS) stimulated CD14+ macrophages alone, 2) co-cultured with CD4+CD45+RO+ or CD4+CD45+RA+ T cells, or 3) anti-CD3 Ab stimulated CD4+CD45+RO+ T cells were incubated in the presence of EGCG (0-10 ng/μl) for 3 and 6 days. Supernatants were collected for cytokine analysis by ELISA. Results: LPS stimulated CD14+ macrophages, or CD4+CD45+RO+ T cells co-cultured with LPS stimulatedmacrophages experienced a dose-dependent reduction in pro-inflammatory cytokine production as shown in table 1. Co-culture of CD14+ macrophages with CD4+CD45+RO+ T cells resulted in a 92% reduction of IL-17 production. There was no change in IL-17 production when CD4+CD45+RA+ T cells were substituted. When evaluated in LPS stimulated CD14+ macrophages alone, EGCG elicited a similar reduction in cytokine production. Reduced cytokine levels were not due to loss of cell viability. Discussion: These results reveal that EGCG produces a significant anti-inflammatory effect by reducing the production of the pro-inflammatory cytokines TNFα, IL-1β, IL-6, and IFNγ. The ability of EGCG to increase apoptosis in lymphocytes from colitis patients, but not from healthy controls, suggests that it may offer a clinical benefit by this mechanism, as well. We are currently conducting a clinical trial in patients with ulcerative colitis to evaluate the clinical correlation of these findings. Table 1