Journal: Scientific Reports

Article Title: Inhibition of corticotropin-releasing hormone receptor 1 and activation of receptor 2 protect against colonic injury and promote epithelium repair

doi: 10.1038/srep46616

Figure Lengend Snippet: Photomicrographs of hematoxylin and eosin (H&E) stained ( A – E ) and immunofluorescence of Mucin 2 (Muc2; mucous-forming protein) ( F – J ) in proximal colon in all experimental groups. Histological scores ( K ) were highest in MS, demonstrated injury in MS compared to control. Treatment with Antalarmin and Astressin prevented this MS-induced colonic injury, but not by Astressin-2β. Crypt length in μm ( L ) (red lines in photomicrographs A – E ) and the number of Muc2+ goblet cells per crypt ( M ) were reduced by MS compared to control, and restored to control levels following Antalarmin and Astressin treatment. Astressin-2β did not prevent these MS-induced effects. Myeloperoxidase (MPO; μmol/mg protein) expression was increased in MS group and was reduced to a level similar to control by treatment with Antalarmin but not by treatment with Astressin or Astressin-2β ( N ). Western blot analysis of NF-κB showed an increase in the phosphorylated expression of NF-κB in MS, which was prevented by Antalarmin administration, but not by Astressin or Astressin-2β ( O , P ). Trans-cellular flux of HRP (ng/ml.cm2.min; Q ) measured by Ussing Chamber was increased in MS and MS + Astressin-2β groups, compared to control, but not in MS + Antalarmin and MS + Astressin groups ( P ). Results are means, ±SD. p < 0.05 was considered significant.

Article Snippet: Staining for Muc2 (NB120-11197, Novus Biologicals, CO) and Ki67 (ab15580, Abcam Inc., Cambridge, MA) was performed to measure epithelial goblet cell numbers and enterocyte proliferation, respectively.

Techniques: Staining, Immunofluorescence, Control, Expressing, Western Blot

Journal: Scientific Reports

Article Title: Inhibition of corticotropin-releasing hormone receptor 1 and activation of receptor 2 protect against colonic injury and promote epithelium repair

doi: 10.1038/srep46616

Figure Lengend Snippet: Fluorescent micrographs of intestinal stem cell marker Lgr5 ( A – E , blue arrows) and cell proliferation marker Ki67 ( F – J ) in colonic tissues of the experimental groups. Lgr5 positive cells expressing Ki67 are shown in higher magnification ( K – N , yellow arrows). Co-localization of Lgr5 and Ki67 markers in the intestine of pups subjected to MS suggested that Lgr5+ intestinal stem cells are proliferative. Relative gene expressions of Lgr5 ( O ) and the number of Ki67+ proliferating cells per crypt (P) in the colon were significantly increased by MS compared to control, and this increase was observed when Antalarmin was administered. In contrast, this increase was prevented by pre-treatment with both Astressin and Astressin-2β. Relative gene expressions of epithelial differentiation marker Muc2 ( Q ) and Lyz1 ( R ) are shown, The MS-induced decrease in Muc2 and Lyz1 expression was rescued by Antalarmin and Astressin, but not Astressin-2β. Relative gene expression IL-22 ( S ) and western blot of phosphorylated STAT3 ( U ) are shown. IL-22 and phosphorylated STAT3 increased in the MS group compared to the control; however, these elevations were both inhibited by Astressin and Astressin-2β, indicating the important role of CRHR2 in tissue repair in response to MS-induced injury. Results are means, ±SD. p < 0.05 was considered significant.

Article Snippet: Staining for Muc2 (NB120-11197, Novus Biologicals, CO) and Ki67 (ab15580, Abcam Inc., Cambridge, MA) was performed to measure epithelial goblet cell numbers and enterocyte proliferation, respectively.

Techniques: Marker, Expressing, Control, Gene Expression, Western Blot