Background Krüppel-like aspect 4 (KLF4) is certainly a zinc finger transcription

Background Krüppel-like aspect 4 (KLF4) is certainly a zinc finger transcription aspect portrayed in the differentiated epithelial cells coating from the intestine. Additionally WT mice provided DSS and nanoparticle/Klf4-siRNA had been less delicate to colitis and acquired reduced Klf4 appearance and while preserving the proliferative response in the colonic epithelium. Conclusions Our outcomes indicate that Klf4 can be an essential mediator of DSS-induced colonic irritation by modulating NF-κB signaling pathway and may be involved in the pathogenesis and/or propagation of inflammatory bowel disease. Thus Klf4 may represent a novel therapeutic target in inflammatory bowel disease. from the intestine have been previously described.36 They have altered differentiation proliferation migration and positioning of intestinal epithelial cells demonstrating an essential role for KLF4 in AM679 maintaining normal intestinal epithelial homeostasis.36 In this study we provide the first evidence that Klf4 in the colonic epithelium plays a crucial role in promoting DSS-induced colitis by modulating NF-κB pathway inflammatory response. MATERIALS AND METHODS Generation of Mice with Intestine-specific Deletion of the Klf4 Gene C57BL/6 mice carrying floxed gene (recombinase gene under the regulation of promoter (in their intestinal epithelium were generated by mating mice with mice followed AM679 by backcrossing to produce mice with intestinal specific deletion of (mutant mice (test and one-way analysis of variance. RESULTS Intestine-specific Deletion of Klf4 Renders Mice Less Susceptible to DSS-induced Colitis To determine the role of deleting Klf4 in DSS-induced colitis mice with or without intestine-specific Klf4 deletion or and mice had no Mouse Monoclonal to HSP60. significant weight change over the experimental period (Fig. 1A). given DSS showed significant weight loss compared with control mice; whereas on the other hand mice showed significantly less weight loss compared with DSS-treated (Fig. 1A). Compared with DSS-treated mice mice had overall significantly lower clinical score and MPO activity (Fig. 1B-E). The protection of mice from DSS-induced colitis was further confirmed by examining H&E-stained colon sections form DSS-treated and mice. As shown in Fig. 2 mice had increased loss of colonic epithelium AM679 (Fig. 2A B) whereas mice had minimal colonic epithelium loss and inflammation(s). FIGURE 1 Resistance of mice to symptoms of DSS-induced colitis. A Mice with intestinal deletion of Klf4 (mice after DSS treatment. A and B H&E staining of DSS-treated mice colon showed extensive colonic epithelium loss. C and D H&E staining of DSS-treated … AM679 Colonic NF-κb Signaling Pathway Is Suppressed After DSS Treatment of Mice with Intestine-specific Deletion of Klf4 (and mice given DSS or not. As shown in Fig. 3A western blot analysis of Klf4 protein level in mice was increased in response to DSS treatment and as expected mice had no or very low levels of Klf4 even after DSS treatment. Relative Klf4 mRNA levels mirrored the change in Klf4 expression level shown in Fig. 3A (see Fig. A Supplemental Digital Content 2 NF-κB has been shown to be activated by DSS treatment43 and to play an important role in intestinal inflammation.44-46 Additionally Klf4 has been shown to mediate NF-κB signaling pathway. 32 33 Consistent with the previous findings mice had low-to-moderate increase of IκB (a suppressor of NF-κB) after DSS treatment whereas mice had relatively higher levels of IκB after DSS treatment as compared with DSS-treated mice (Fig. 3A). AM679 Staining for NF-κB (p65 subunit) showed basal nuclear localization and comparable staining level of NF-κB in the colonic epithelium in both and mice (Fig. 3B 1 and 2 respectively). However after DSS treatment mice had increased cytoplasmic and nuclear staining AM679 of NF-κB (Fig. 3B 3 as compared with untreated mice. Interestingly mice showed reduction both in overall staining and in the nuclear localization of NF-κB after DSS treatment (Fig. 3B 4 as compared with both untreated and mice. On analyzing the mRNA levels of inflammatory cytokines Il-1β Il-6 and TNFα after DSS treatment mice had significantly lower levels of these cytokines when compared with mice (see Fig. B-D Supplemental Digital Content 2 Additionally quantitation of inflammatory cell infiltrates: macrophages (F4/80) lymphocytes (CD11c) granulocytes and monocytes (Ly-6G) and for.