Vol. 305, Issue 1, 232-239, April 2003

Significant Role of Ceramide Pathway in Experimental Gastric Ulcer Formation in Rats

Keita Uehara, Soichiro Miura, Tetsu Takeuchi, Takao Taki, Manabu Nakashita, Masayuki Adachi, Toshiaki Inamura, Toshiko Ogawa, Yasutada Akiba, Hidekazu Suzuki, Hiroshi Nagata and Hiromasa Ishii

Department of Internal Medicine, School of Medicine, Keio University, Tokyo, Japan (K.U., Te.T., M.N., M.A., T.I., T.O., Y.A., H.S., H.N., H.I.); Second Department of Internal Medicine, National Defense Medical College, Saitama, Japan (S.M.); and Molecular Medical Science Institute, Otsuka Pharmaceutical Co., Ltd., Tokushima, Japan (Ta.T.)

Ceramides have emerged as key participants in the signaling pathway of cytokines and apoptosis. We previously revealed that phorbol 12-myristate 13-acetate (PMA) induced experimental ulcers in rat gastric mucosa. In this study, we investigated the role of ceramide in ulcer formation and its relation to the activation of transcription factors and apoptosis. PMA was subserosally injected to rat glandular stomach. Fumonisin B1 (FB1), an inhibitor of ceramide synthase, was administered together with the PMA. The time course of ceramide content was quantified using thin layer chromatography and the number of apoptotic cells was determined by immunohistochemistry. The activation of transcription factor nuclear factor-B (NF-B) or activator protein-1 (AP-1) was evaluated using an electrophoretic mobility shift assay. The administration of FB1 attenuated PMA-induced gastric ulcer formation in a dose-dependent manner. Before the ulcers became obvious, the ceramide content (C18 and C24 ceramide) increased significantly in the gastric wall. The activation of NF-B and AP-1 and an increase in the number of apoptotic cells were also observed. Both of these were significantly inhibited by the coadministration of FB1. However, NF-B inhibitors attenuated gastric ulcer formation without affecting the ceramide content or the number of apoptotic cells. Ceramide formation in the stomach significantly contributes to PMA-induced tissue damage, possibly via the activation of transcription factors and an increase in apoptosis in the gastric mucosa. However, after the increase in ceramide levels, the NF-B and apoptosis pathways may be separately involved in ulcer formation.