Chios Mastic Fractions in Experimental Colitis: Implication of the Nuclear Factor κB Pathway in Cultured HT29 Cells

Abstract

The Pistacia lentiscus tree gives a resinous exudate called Chios mastic (CM) rich in triterpenoids. CM can be fractionated into acidic and neutral fractions (AF and NF, respectively). Oleanolic acid (OA) is a major triterpenic acid in CM with several antioxidant and anti-inflammatory properties. We have recently shown that CM is beneficial in experimental colitis in the form of powder mixture with inulin, as supplied commercially. However, the bioactive fraction or compound of CM is unidentified. Thus, based on the hypothesis that terpenoids exhibit functional activities via distinguishable pathways, we fractionated CM and applied different fractions or individual OA in experimental colitis. Furthermore, we investigated the mechanism underlying this effect in human colon epithelial cells. CM powder mixture (100 mg/kg of body weight) or the respective CM powder mixture components (i.e., inulin, AF, NF, or OA) were individually administered in trinitrobenzene sulfonic acid-treated rats. Colonic damage was assessed microscopically, and levels of tumor necrosis factor-α (TNF-α), interleukin (IL)-6, IL-8, and intercellular adhesion molecule-1were measured. A model of inflammation in co-cultured human colon epithelial HT29 cells and monocytes/macrophages was established. Lactate dehydrogenase release and levels of TNF-α, IL-8, and nuclear factor-κB (NF-κB) p65 were measured. In vivo, histological amelioration of colitis and significant regulation in inflammation occurred with CM powder mixture, even at the mRNA level. Although no histological improvement was observed, AF and NF reduced levels of inflammatory markers. Inulin was ineffective. In vitro, CM treatment down-regulated IL-8 and NF-κB p65. Neither fractions nor OA was the bioactive component solely. Most probably, the entire CM rather than its individual fractions reduces inflammation via NF-κB regulation.