T1158 Ameliorating Intestinal Inflammation and Preventing Colon Tumor Development by the Plant Extract, Berberine

Abstract

Background. Berberine, an isoquinoline alkaloid derived from plants, has been used as a traditional medicine for several diseases, including bacterial diarrhea. Recently, Berberine has been shown to suppress tumor cell growth and regulate several signaling pathways involved in apoptosis and cell cycle arrest In Vitro. However information is limited regarding mechanisms of Berberine's action in diseases. This study was designed to investigate Berberine's anti-tumor and anti-inflammatory effects. Methods. 7 patients with familial adenomatous polyposis history and recurrent colon adenomatous polyps were enrolled for oral Berberine treatment (0.1 g, 3 times/day) immediately following polypectomy and patients followed-up up to two years. Animal studies were performed on C57BL/6 mice administered 3% DSS in drinking water, or water only, for 7 days. Mice were given Berberine (200 μg/ g body weight, once daily) via gavage from day 4-7. Colon sections were prepared for H&E staining to determine intestinal tissue injury score (0: normal, 18: severe colitis). Mouse colon epithelial cells derived from wt Immorto (YAMC), Immorto-Min mice carrying APC min mutation (IMCE), and IMCE cells over-expression of Ras (IRas) were treated with Berberine (25 μM 200 μM) in the presence or absence of TNF (100 ng/ml), EGF (10 ng/ ml) or caspase inhibitor for detecting apoptosis using Annexin staining and TUNEL assay, signaling by Western blot analysis, and colony formation in agar. Results. 6 (7 total) patients showed no recurrence of polyps during the 2-year Berberine treatment. Berberine induced apoptosis in IMCE and IRas, but not YAMC cells and inhibited IRas cell colony formation in a concentration-dependent manner. Berberine stimulated p53 activation and blocked TNF-induced NFκB activation. In addition, Berberine inhibited EGF-stimulated EGF receptor activation and cell proliferation. However, Berbeine did not activate caspase 3 or 9 nor did the caspase inhibitor prevent Berberine-induced apoptosis. DSS-induced colitis (score: 15±0.9) was significantly reduced by Berberine treatment (score: 10±1.3, p<0.05). Conclusion. Berberine exerts the anti-tumor effect, which may be through induction of tumor cell apoptosis and inhibition of the action of tumor-promoting factors, including EGF and TNF. In addition, anti-inflammatory effects of Berberine may contribute to prevention of colitisassociated tumor development. The role of p53 activation and NFκB inhibition by Berberine in prevention of tumor development and inflammation is under investigation. Thus, Berberine may serve as a potential alternative approach for treating intestinal inflammatory diseases and tumor development.