cyclic-gmp and Esophagitis

We investigated whether the signal mechanism for relaxation may be affected by inflammation of the cat esophagus. Acute esophagitis was induced by perfusion with 0.1N HCI at a rate of 1 mL/min for 45 min over three consecutive days. We then isolated esophageal smooth muscle cells by enzymatic digestion with collagenase. We pre-contracted the isolated smooth cells with acetylcholine (ACh) (10(-5) M) and compared the agonist-induced relaxation of pre-con tracted normal cells with those of esophagitic cells. Vasoactive intestinal polypeptide (VIP) caused a dose-dependent relaxation in normal cells, and this curve was down shifted in esophagitic cells. Sodium nitroprusside (SNP) or SIN-1 (NO donor) produced dose-dependent relaxation in normal cells, which was not affected by esophagitis. 8-Br-cGMP (a cGMP ana log) also induced dose-dependent relaxation to a similar extent in both normal and esoph agitic cells. Forskolin (a cAMP activator) or db-cAMP (a cAMP analog) produced dose-dependent relaxation in normal cells, and this relaxation curve was down shifted in esoph agitic cells. Western blotting was used to determine what subtype of adenylyl cyclase was involved in the cAMP pathway. Western blot analysis of homogenates derived from esophageal smooth muscle using antibodies against adenylyl cyclase types II, III, IV and V/VI revealed the presence of type V and/or type VI only. This result suggests that relaxation via a cAMP-dependent pathway rather than a cGMP dependent-pathway is down regulated in cat acute esophagitis. This subsensitivity of the cAMP related pathway may be related to the activ ity of adenylyl cyclase V/VI.

Topics: Adenylyl Cyclases; Animals; Blotting, Western; Bucladesine; Cats; Cell Membrane Permeability; Colforsin; Cyclic AMP; Cyclic GMP; Dose-Response Relationship, Drug; Down-Regulation; Esophagitis; In Vitro Techniques; Isoenzymes; Molsidomine; Muscle Relaxation; Muscle, Smooth; Nitric Oxide Donors; Nitroprusside; Signal Transduction; Vasoactive Intestinal Peptide

The present study explores the changes of nitric oxide synthesis and esophageal dysmotility in a feline model of esophagitis. Perfusion of the esophagus with acid produced inflammatory changes of esophageal mucosa. The esophageal motility was measured before and after the perfusion. The nitric oxide synthase activity, the l-arginine uptake, and the content of cyclic guanine monophosphate of the muscle and the mucous membrane were determined and the NADPH-diaphorase was stained. Esophagitis impairs the motility of the esophagus. The nitric oxide synthase activity, the content of cyclic guanine monophosphate, the NADPH-diaphorase stain and the maximum velocity of l-arginine uptake of lower esophageal sphincter of the cats in the acid perfusion group were higher than those of the control group. The maximum velocity of l-arginine transport and the content of cyclic guanine monophosphate of the mucosa in the acid perfusion group were lower than those of the control group. The results suggested that during esophagitis there is an alteration of the l-arginine/nitric oxide synthase/nitric oxide pathway in the esophagus, which may be one of the important mechanisms of esophageal motility dysfunction.

Topics: Animals; Arginine; Cyclic GMP; Disease Models, Animal; Esophageal Motility Disorders; Esophagitis; Male; Nitric Oxide; Nitric Oxide Synthase; Probability; Random Allocation; Reference Values; Sensitivity and Specificity

It has been previously shown that induction of experimental esophagitis in the cat by esophageal perfusion for 30 minutes with 0.1N HCl for 4 consecutive days results in a significant reduction of in vivo lower esophageal sphincter (LES) resting pressure and in vitro spontaneous tone without affecting esophageal response to KCl. It has also been shown that basal LES tone and LES contraction in response to acetylcholine depend on the release of calcium from intercellular stores, whereas esophageal contraction is mediated by extracellular calcium. The present report shows that esophageal acid perfusion impairs the transduction pathway mediating lower esophageal sphincter contraction in response to acetylcholine through release of intracellular calcium because LES strips and single cells no longer contract in response to acetylcholine if calcium is removed from the physiologic salt solution. This suggests that either the intracellular calcium stores or the release mechanisms that mediate maintenance of tone and contraction in response to acetylcholine may be damaged. However, the acid perfusion has no effect on the acetylcholine response in the esophagus, which is mediated by the influx of extracellular calcium. In the LES circular muscle, the injury results in reduced levels of inositol phosphates without affecting resting levels of 5'-cyclic adenosine monophosphate or 5'-cyclic guanosine monophosphate. The reduced levels of 1,4,5-inositol trisphosphate are consistent with impairment in the mechanisms responsible for release of intracellular calcium, although concurrent damage to calcium stores may also occur.

Topics: Acute Disease; Animals; Calcium; Cats; Cyclic AMP; Cyclic GMP; Egtazic Acid; Esophagitis; Esophagogastric Junction; Female; In Vitro Techniques; Inositol Phosphates; Male; Signal Transduction