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The effect of sulphasalazine and 5-aminosalicylic acid on the human colon myofibroblasts

Jolanta Lodowska 2Arkadiusz R. Gruchlik 1Daniel Wolny 1Joanna M. Wawszczyk 2Zofia Dzierżewicz 1Ludmiła Węglarz 2

1. Department of Biopharmacy, Medical University of Silesia, Jedności 8, Sosnowiec 41-200, Poland
2. Department of Biochemistry, Medical University of Silesia, Jedności 8, Sosnowiec 41-200, Poland


Sulfasalazine (SAS) is a drug commonly used in the treatment of inflammatory intestinal diseases. In addition to its bacteriostatic, anti-inflammatory and immunosuppressive action, it greatly reduces the risk of neoplastic lesions of the colon and rectum in patients suffering from ulcerative colitis. Sulfasalazine therapeutic effect results from pharmacological activity of 5-aminosalicylic acid (ASA), the metabolite of SAS formed in the reaction catalysed by bacterial azo-reductases. The number of myofibroblasts, the cells involved in organogenesis, growth and differentiation of the intestinal epithelial cells, mucosa regeneration as well as in fibrosis and carcinogenesis process, significantly increases in the course of inflammatory bowel diseases.

The aim of the study was to evaluate the effect of SAS and its active metabolite (ASA) on the viability of colon subepithelial myofibroblasts.

The tests were performed on normal human myofibroblasts CCD-18Co derived from the American Type Culture Collection (ATCC, Manassas, VA). Cells were cultured in Minimal Essential Medium supplemented with 10% fetal bovine serum (FBS), 100 IU/ml penicillin G and 100 mg/ml streptomycin (Gibco) and 10 mM HEPES. Cell were cultured at 37°C in a 5% CO2 atmosphere. Cytotoxicity of SAS and ASA against myofibroblasts was evaluated with XTT (Toxicology In Vitro Test Kit, XTT based, TOX-2, Sigma). Cells were seeded on 96-well plate at density of 3000 cells/well. Subsequently, they were cultured in the presence of 0.5 mM, 1 mM, 2.5 mM, 5 mM and 10 mM ASA, and 0.05 mM, 0.1 mM, 0.25 mM, 0.5 mM and 1 mM SAS for 72 hours at standard conditions. Myofibroblasts were incubated with XTT solution for 4 hours and absorbance was measured at 450 nm (reference 690 nm) using a plate reader.

It was found that 1 mM SAS and 5 mM ASA decreased the number of myofibroblasts as compared to the control, which could be explained by the cytotoxic action of these compounds against investigated cells.


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Submitted: 2014-03-13 11:39
Revised:   2014-05-02 19:41