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inhibits intestinal smooth muscle proliferation in an organ culture system: Involvement of COX-2 and iNOS induction in muscularis resident macrophages
1 Veterinary Pharmacology, Univ of Tokyo, Tokyo, Japan
2 Pathology, NIAH, Tsukuba, Japan
3 Pharmacology, Univ of Nevada Reno, Reno, Nevada, United States
* To whom correspondence should be addressed. E-mail: aozaki{at}mail.ecc.u-tokyo.ac.jp.
Intestinal inflammation causes hyperplasia of smooth muscle that leads to thickening of the smooth muscle layer, resulting in dysmotility. IL-1
is a pro-inflammatory cytokine that plays a central role in intestinal inflammation. In this study, in order to evaluate the effect of IL-1 on proliferation of ileal smooth muscle cells in in vivo, we utilized an organ culture system. When rat ileal smooth muscle tissue was cultured under serum free conditions for 3 days, most smooth muscle cells were maintained their arrangement and kept their contractile phenotype. When 10% fetal bovine albumin (FBS) was added, increased number of smooth muscle cells per unit area was observed. Moreover, immunohistochemical staining for PCNA demonstrated that FBS induced proliferation of smooth muscle cells. IL-1 inhibited the proliferative effect of FBS. Furthermore, IL-1 up-regulated iNOS and COX-2 mRNA and protein, and thus stimulated NO and PGE2 productions. Moreover, exogenously applied NO and PGE2 inhibited the increase of BrdU positive cells stimulated with FBS. Immunostaining revealed that the majority of COX-2 and iNOS was located in the dense network of macrophages resident in the muscularis which were immuno-reactive to ED2. Based on these findings, IL-1 acts as an anti-proliferative mediator, which acts indirectly through the production of PGE2 and NO from resident macrophage within ileal smooth muscle tissue.
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