Real-time quantitative PCR analysis of mouse colon samples

Gut mucosal healing (MH) is considered a key therapeutic target and prognostic parameter in the management of inflammatory bowel disease (IBD). The dipotassium glycyrrhizate (DPG), a salt of the glycoconjugated triterpene glycyrrhizin, has been shown to inhibit the High Mobility Group Box 1 (HMGB1) protein, an allarmin strongly implicated in the pathogenesis of most inflammatory and auto-immune disorders, included IBD. The aims of the present study is to identify genes involved in MH pathways modulated by DPG in mouse model of chemical induced acute colitis. To induce colitis C57BL/6 female mice were treated 3% dextran sodium sulphate (DSS) for 7 days and then sacrificed. Mice, were randomly divided into three groups: 1) control group received regular drinking water; 2) mice treated with 3% DSS; 3) mice treated with 3% DSS and 8 mg/kg/day DPG. RNA obtained from colon tissue of controls (Untreated), DSS-induced colitis and mice co-treated with DSS and DPG, were analyzed by RT2 PCR array. PCR-Array was repeated in triplicate using a pool of RNA obtained from 5 animals for each group.