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|Title:||Regulation of Sodium Hydrogen Exchanger Isoform-2 in Experimental Colitis||Authors:||Amal Ali Hassan Sulaiman||Supervisor:||Prof. Islam Khan||Keywords:||Fulfillment||Issue Date:||2017||Publisher:||Kuwait university - college of graduate studies||Abstract:||The Na+/H+ exchanger (NHE) belongs to a ubiquitous family of transmembrane proteins which is represented by a group of 10 distinct genes (NHE-1-10). They mediate counter-transport of extracellular Na+ for intracellular H+ and play an important role in the regulation of intracellular pH and cell volume. Altered NHE activity has been linked to the pathogenesis of several diseases including inflammatory bowel diseases (IBD). In this study, we examined the role of NHE-2 isoform in experimental colitis. This study employed Sprague-Dawley male rats. Animals were divided into two groups; the non-colitis controls and the colitis. Colitis was induced by delivering 30 mg of 2,4,6-trinitrobenzenesulfonic acid (TNBS) solution in 50% ethanol intra-rectally in each rat. The animals were sacrificed by cervical dislocation on the day 6 post-TNBS treatment. There was a significant decrease in the body weight of the colitis animals over the test period. Histological evaluation showed depletion of goblet cells, muscle thickening, decreased mucin level and increase in MPO activity in inflamed rat colon. NHE-2 protein expression levels were decreased in inflamed colon as measured by western blot analysis whereas actin, an internal control remained unchanged. Confocal immunofluorescence microscopy also showed a significant decrease in the level of NHE-2 protein in inflamed colon. Furthermore, NHE-2 isoform was co-localized with the NHE-3 apical isoform on the brush border of epithelial cells in the colon. We used an end-point RT-PCR method for quantitation of NHE-2 mRNA, and the data was confirmed with SYBR green real-time PCR. Both methods showed a significant decrease in the level of NHE-2 mRNA expression in the inflamed colon. To further investigate the mechanism of mRNA reduction, the level of NHE-2 HnRNA was measured and found to be significantly reduced in the inflamed colon. A similar reduction in both mRNA and HnRNA suggests that the decrease in protein is regulated transcriptionally. Suppression of NHE-2 is expected to disturb electrolyte balance, and hence may contribute in part to the symptoms of inflammatory bowel diseases such as diarrhea||URI:||http://hdl.handle.net/123456789/626|
|Appears in Programs:||0540 Medical Biochemistry (M.Sc.)|
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