Evaluation of DA-6034 for treatment effect of ulcerative colitis:Analysis of transcriptome and proteome profiles

Abstract

Backgrounds; Inflammatory bowel disease(IBD) is a multifunctional disorder with unknown etiology and pathogenesis, resulting in no specific treatment modality until now. The effect of DA-6034, a new flavonoid derivative showing favorable effects in animal models of IBD, on gene expression patterns at the lesion site was investigated in experimental colitis using cDNA microarray and proteomic analysis with SELDI-TOF to explore the underlying pathogenesis and the mode of action of the potential drug. Methods; Experimental colitis was induced by intracolonic administration of 30 mg of TNBS in 0.5ml of 50% ethanol to fasted albino rats. Then, DA-6034(0.3 to 10 mg/kg) was treated via enema once daily for 7 days starting from day 1 after the injury. At 24 hours after the last drug treatment, gross lesion and histological scoring was conducted. For proteomic analysis, 2D gel electrophoresis and SELDI-TOF analysis were performed. In addition, cDNA microarray and real time PCR were conducted to match the genes identified from proteomic analysis before and after DA-6034 treatment. Results; DA-6034 reduced gross lesion score and malondialdehyde levels in TNBS-induced colitis siginificantly. cDNA microarray revealed that more than 80 genes are siginificantly upregulated before and after DA-6034 treatment. Some of them were identified as genes involved in lymphocytic homing and inflammation associated genes. We also found 10 proteins are significantly decreased or increased in experimental colitis like S100 calcium binding protein A6(calcyclin), nuclear protein stromal antigen 1, DKFZP434F091 protein, and 6-PF-2-kinase-2,6-bisphosphatase 4. Conclusion; These results suggest the genome analysis using high throughput techniques can enable identification of the candidate genes involved in the generation of ulcerative colitis and the possible target of candidate drugs.