Basic Research
Copyright ©2005 Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Mar 14, 2005; 11(10): 1508-1514
Published online Mar 14, 2005. doi: 10.3748/wjg.v11.i10.1508
Activation of nuclear factor-kappa B and effects of pyrrolidine dithiocarbamate on TNBS-induced rat colitis
Ken Chen, You-Ming Long, Hui Wang, Lei Lan, Zhen-He Lin
Ken Chen, You-Ming Long, Department of Clinical Medicine, Guangdong College of Pharmacy, Guangzhou 510224, Guangdong Province, China
Hui Wang, Department of Pharmacology, Guangdong Medical College, Zhanjiang 524023, Guangdong Province, China
Lei Lan, Zhen-He Lin, Department of Digestion, the Affiliated Hospital of Guangdong Medical College, Zhanjiang 524023, Guangdong Province, China
Author contributions: All authors contributed equally to the work.
Supported by a Grant From Health Department Foundation of Guangdong Province, No. A2003554
Correspondence to: Professor Ken Chen, Department of Clinical Medicine, Guangdong College of Pharmacy, Guangzhou 510224, Guangdong Province, China. chken@gdpc.edu.cn
Telephone: +86-20-34074077
Received: June 30, 2004
Revised: July 1, 2004
Accepted: July 11, 2004
Published online: March 14, 2005
Abstract

AIM: To explore the changes of nuclear factor-kappa B (NF-κB) DNA-binding activity, the expression of intercellular adhesion molecule-1 (ICAM-1) regulated by NF-κB at various times and to evaluate the effects of pyrrolidine dithiocarbamate (PDTC) on trinitrobenzene sulfonic acid (TNBS)-induced rat colitis.

METHODS: TNBS of 0.6 mL was mixed with ethanol of 0.3 mL solution and instilled into the lumen of the rat colon. The rat models were divided into 6 groups, which were killed at 24 h, 3, 7, 14, and 21 d after enema. Colonic inflammation and damage were assessed by macroscopical and histological criteria. Activity of NF-κB DNA-binding was analyzed by electrophoresis mobility shift assays (EMSA). Expression of ICAM-1 was detected by in situ hybridization (ISH) and immunohistochemistry (IH). Then various doses of PDTC were injected into rat abdomen 30 min before enema with TNBS/ethanol as pretreatment. The rats were killed 4 h after enema and the colonic inflammation, myeloperoxidase (MPO) activity, malondialdehyde (MDA) level, and DNA-binding activity of NF-κB were assessed. Finally, PDTC was injected intraperitoneally after colitis was induced. Changes of morphology were assayed.

RESULTS: During the first week, hyperemia, hemorrhage, edema and ulceration of the colonic mucosa appeared with predominant infiltration of leukocytes. Neutrophils, macrophages, lymphocytes infiltrated in mucosa and submucosa 14 d later. Fibroblasts and granuloma-like structures were also obviously seen. The binding activity of NF-κB began to increase at 24 h time point and reached a peak at 14 d, then decreased but still was higher than control group at 21 d (P<0.01). Levels of ICAM-1 mRNA and protein significantly elevated at 24 h and the peak was at 21 d. Pretreatment with PDTC could attenuate the development of inflammation but not by reducing NF-κB activity. This attenuation of inflammation had a positive relationship with the dose of PDTC. PDTC at the dose of 100 mg/kg had no therapeutic effect after colitis was induced.

CONCLUSION: NF-κB activation is an important event that may be involved in acute and chronic inflammation development and may contribute to self-protection against early inflammation damage. NF-κB also regulates ICAM-1 expression during colonic inflammation. Pretreatment of PDTC may attenuate the inflammation development. But PDTC has no therapeutic effect after the colitis is induced.

Keywords: Nuclear factor; Pyrrolidine dithiocarbamate; Rat; Colitis