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Am J Physiol Gastrointest Liver Physiol 275: G68-G75, 1998;
0193-1857/98 $5.00
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Vol. 275, Issue 1, G68-G75, July 1998

Substance P receptor expression in intestinal epithelium in Clostridium difficile toxin A enteritis in rats

Charalabos Pothoulakis1, Ignazio Castagliuolo1, S. E. Leeman2, Chi-Chung Wang1, Hanzong Li2, Beth J. Hoffman3, and Eva Mezey3

1 Division of Gastroenterology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston 02215; 2 Department of Pharmacology, Boston University School of Medicine, Boston, Massachusetts 02118; and 3 Laboratory of Cell Biology, National Institute of Mental Health, Bethesda, Maryland 20892

We previously reported that the inflammatory effects of Clostridium difficile toxin A on rat intestine can be significantly inhibited with a specific neurokinin-1 receptor (NK-1R) antagonist. In this study we investigated the localization and expression of NK-1R mRNA and protein in rat intestine by in situ hybridization, Northern blot analysis, and immunohistochemistry, respectively, after exposure to toxin A. Northern blot analysis showed increased mucosal levels of NK-1R mRNA starting 30 min after toxin A administration. In situ hybridization showed that toxin A increased NK-1R mRNA expression in intestinal epithelial cells after 30, 120, and 180 min. In rats pretreated with the NK-1R antagonist CP-96345 the increase in NK-1R mRNA levels after exposure to toxin A was inhibited, indicating that NK-1R upregulation is substance P (SP) dependent. One hour after exposure to toxin A many of the intestinal epithelial cells showed staining for NK-1R compared with controls. Specific I-SP binding to purified epithelial cell membranes obtained from ileum exposed to toxin A for 15 min was increased twofold over control and persisted for 4 h. This report provides evidence that NK-1R expression is increased in the intestinal epithelium shortly after exposure to toxin A and may be important in toxin A-induced inflammation.

intestinal inflammation; neuropeptides; neurokinin-1 receptor


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