Paul H. Guth

Microcirculatory stasis precedes tissue necrosis in ethanol-induced gastric mucosal injury in the rat

The relation of blood flow stasis to the development of unequivocal histologic necrosis (loss of parietal cells from the column of contiguous cells) in ethanol-induced gastric mucosal injury was studied in anesthetized rats. The most rapid vascular change that occurred when the gastric mucosa was exposed to 100% ethanol was a severe segmental constriction of the large submucosal venules. At 22 sec, the average venular diameter was 52.2±6.0% of the original one. This was followed by complete superficial mucosal blood flow stasis at 49±4 sec and appearance of histologic evidence of necrosis in one of seven rats at 2.5 min, four of six rats at 10 min, and seven of seven rats at 60 min. We conclude that in ethanol-induced gastric mucosal injury, submucosal venular constriction occurs first, followed by cessation of mucosal blood flow to be followed later on with histologic evidence of necrosis.

Acid-sensing pathways of rat duodenum

We tested the hypothesis that the duodenal hyperemic response to acid occurs through activation of capsaicin-sensitive afferent nerves with subsequent release of vasodilatory substances such as calcitonin gene-related peptide (CGRP) and nitric oxide (NO). Laser-Doppler flowmetry was used to measure duodenal blood flow in urethan-anesthetized rats. Duodenal mucosa was superfused with pH 7. 0 buffer with capsaicin or bradykinin or was acid challenged with pH 2.2 solution, with or without vanilloid receptor antagonists, a CGRP receptor antagonist, an NO synthase (NOS) inhibitor, or a cyclooxygenase inhibitor. The selective vanilloid receptor antagonist capsazepine (CPZ) dose dependently inhibited the hyperemic response to acid and capsaicin but did not affect bradykinin-induced hyperemia. Ruthenium red was less inhibitory than capsazepine. Selective ablation of capsaicin-sensitive nerves, CGRP-(8-37), and N(G)-nitro-L-arginine methyl ester inhibited acid-induced hyperemia, but indomethacin did not. We conclude that luminal acid, but not bradykinin, stimulates CPZ-sensitive receptors on capsaicin-sensitive afferent nerves of rat duodenum. Activation of these receptors produces vasodilation via the CGRP-NO pathway but not via the cyclooxygenase pathway. Acid appears to be the endogenous ligand for duodenal vanilloid receptors.We tested the hypothesis that the duodenal hyperemic response to acid occurs through activation of capsaicin-sensitive afferent nerves with subsequent release of vasodilatory substances such as calcitonin gene-related peptide (CGRP) and nitric oxide (NO). Laser-Doppler flowmetry was used to measure duodenal blood flow in urethan-anesthetized rats. Duodenal mucosa was superfused with pH 7.0 buffer with capsaicin or bradykinin or was acid challenged with pH 2.2 solution, with or without vanilloid receptor antagonists, a CGRP receptor antagonist, an NO synthase (NOS) inhibitor, or a cyclooxygenase inhibitor. The selective vanilloid receptor antagonist capsazepine (CPZ) dose dependently inhibited the hyperemic response to acid and capsaicin but did not affect bradykinin-induced hyperemia. Ruthenium red was less inhibitory than capsazepine. Selective ablation of capsaicin-sensitive nerves, CGRP-(8-37), and N G-nitro-l-arginine methyl ester inhibited acid-induced hyperemia, but indomethacin did not. We conclude that luminal acid, but not bradykinin, stimulates CPZ-sensitive receptors on capsaicin-sensitive afferent nerves of rat duodenum. Activation of these receptors produces vasodilation via the CGRP-NO pathway but not via the cyclooxygenase pathway. Acid appears to be the endogenous ligand for duodenal vanilloid receptors.

Central nervous system action of thyrotropin-releasing hormone to increase gastric mucosal blood flow in the rat

The central nervous system effects of thyrotropin-releasing hormone (TRH) on gastric acid secretion and mucosal blood flow were studied in rats. Corpus mucosal blood flow was measured by the hydrogen gas clearance technique and acid output by a continuous gastric perfusion method in fasted, urethane-anesthetized rats. Thyrotropin-releasing hormone (1 or 5 micrograms) injected into the cerebral lateral ventricle induced concomitant increases in gastric acid secretion and mucosal blood flow. Intravenous infusion of step doses of TRH (60 and 180 micrograms/kg.h) had no effect on these parameters. Bilateral vagotomy and atropine (0.15 mg/kg) completely blocked the effects of intracerebroventricular injection of TRH (5 micrograms) on gastric acid secretion and mucosal blood flow. In contrast, intravenous omeprazole (20 mumol/kg) completely inhibited the increase in gastric acid secretion but not the increase in mucosal blood flow elicited by intracerebroventricular administration of TRH (5 micrograms). These results demonstrate that TRH acts in the brain to stimulate gastric acid secretion and mucosal blood flow through vagal dependent pathways and peripheral muscarinic receptors. Part of the effect of central TRH on gastric mucosal blood flow is not secondary to the stimulation of acid secretion and appears to represent a direct cholinergic vasodilatory response.

Assessment of mucosal hemodynamics in normal human colon and patients with inflammatory bowel disease

Reflectance spectrophotometry measures indices of mucosal hemoglobin concentration (IHB) and oxygen saturation (ISO2). In the rat colon, characteristic patterns of IHB and ISO2 are associated with ischemia with congestion (increased IHB and decreased ISO2) and ischemia without congestion (decreased IHB and decreased ISO2). Endoscopic measurements with acceptable interobserver variability was demonstrated in the canine stomach. In eight healthy subjects, endoscopic measurement in different areas of the colon and rectum revealed significantly lower IHB values in the splenic flexure. These observations are compatible with reduced flow and increased susceptibility to ischemic damage in this watershed area. The endoscopic measurements in 13 patients with active inflammatory bowel disease revealed an increase in IHB and ISO2 values in the involved areas, indicating an increase in mucosal blood flow. In six patients restudied when the disease remitted, these values returned to normal.

Ethanol-induced gastric mucosal blood flow and vascular permeability changes in the rat

Ethanol-induced gastric mucosal injury is accompanied by complete cessation of blood flow in the lesion area. An in vivomicroscopy technique was used in the rat to determine whether this cessation of blood flow could be due to increased microvascular permeability with marked plasma exudation and a resultant increase in blood viscosity. The topical application of 100% ethanol to the mucosa caused complete stasis of mucosal blood flow within 1 min. Fluorescent in vivomicroscopy revealed that topical ethanol also caused a prompt increase in mucosal microvascular permeability to albumin. This, however, did not explain the slowing and cessation of mucosal microvascular blood flow as these preceded the permeability change.

Effect of Nω-nitro-L-arginine methyl ester on arterial pressure and on vasodilator and vasoconstrictor responses: influence of initial vascular tone

Intravenous N omega-nitro-L-arginine methyl ester (L-NAME) 0.3, 3 and 30 mg/kg produced a dose-dependent increase of blood pressure in urethane-anesthetized rats. Similar pressure responses occurred in rats after ganglionic or adrenergic blockade, hemorrhage or acetylcholine infusion. L-NAME potentiated the pressor response to phenylephrine in pentolinium-treated rats. L-NAME increased the depressor effect of acetylcholine and sodium nitroprusside and shortened the acetylcholine response. Phenylephrine potentiated the magnitude but not the duration of the acetylcholine response and had no effect on the duration or magnitude of sodium nitroprusside-induced hypotension. L-NAME potentiated the initial fall of blood pressure induced by a 5 min acetylcholine infusion but had no effect on the pressure drop at the end of infusion. These results suggest that basal production of nitric oxide (NO) is not related to vascular tone and do not support the view that acetylcholine releases NO from resistance vessels in vivo.

Role of oxyradicals in cold water immersion restraint-induced gastric mucosal injury in the rat

Cold water immersion restraint of the rat results in focal gastric mucosal erosions. The lesions are associated with powerful, prolonged-duration gastric contractions. Phasic gastric contractions may attenuate gastric mucosal blood flow, resulting in ischemia followed by reperfusion. Therefore, the conditions of cold-water-immersion restraint might lead to mucosal injury by an oxyradical-mediated mechanism. To test this hypothesis, we studied the effect of oxyradical inhibition on cold water immersion restraint-induced lesions. In separate groups of rats subjected to cold water immersion restraint (6–10 animals per group), oxyradical inhibition was achieved by chronic feeding of a sodium tungstate diet, oral administration of allopurinol, or intraperitoneal administration of dimethylsulfoxide. None of these regimens significantly attenuated the number of lesions per stomach, the total lesion area, or the percent of corpus mucosa containing lesions. We conclude that oxyradicals do not play a role in the pathogenesis of cold water immersion restraint-induced lesions.

Cimetidine and ranitidine protect against cold restraint-induced ulceration in rat by suppressing gastric acid secretion

The effect of the H2-receptor antogonists cimetidine and ranitidine on pentagastrinstimulated gastric acid secretion in anesthetized rats, and gastric mucosal lesion formation and gastric motility in unanesthetized cold-restrained rats was studied. Both cimetidine and ranitidine suppressed pentagastrin-stimulated gastric secretion in a dose-dependent fashion. Cold restraint-induced lesion formation was not prevented with doses of both agents that inhibited acid secretion by 75%. Doses which suppressed pentagastrin-stimulated acid secretion more than 90% significantly prevented the development of gastric mucosal lesions produced by cold restraint. Doses of both H2 blockers which demonstrated significant suppression of lesion formation had no effect on cold restraint-stimulated gastric contractility. We conclude that cimetidine and ranitidine suppress cold restraint-induced lesion formation by suppressing acid secretion and not by suppressing gastric contractility.

Spectrum of injury produced in the duodenum by perfusion with luminal acid in the rat.

The dose and time dependence of duodenal mucosal injury by luminal acid perfusion was studied. Saline, 0.01, 0.05, 0.15, and 0.3N HCl, were perfused through the proximal duodena of rats for 5, 15, or 30 minutes and then harvested for histological examination. In a second set of studies, after a 30-minute perfusion, duodena were harvested either immediately or 2, 4, 8, or 24 hours after the perfusion to study the recovery from injury. Acid disappearance (acid delivered minus acid recovered) was measured in all groups. Duodena were examined grossly, then fixed, stained, and scored histologically. Whereas no gross mucosal injury was noted, there was graded histological injury proportional to acid concentration. Injury occurred early in the perfusion and changed little with increased perfusion durations. The initial injury lead to an acid disappearance rate that was proportional to acid concentration and, therefore, the degree of injury. After the initial injury occurred, the rate of acid neutralization was unchanged by increased duration of acid perfusion. This acid neutralization protected against further injury despite the continued presence of acid. Recovery from injury was complete with physiological (0.01 and 0.05N HCl) but not pharmacological (0.15 and 0.3N HCl) concentrations of acid. It is concluded that acid-induced duodenal injury occurs within 5 minutes of exposure, is proportional to the acid concentration, and results in acid neutralization that protects against extension of the injury with continued acid exposure.

Reduction in index of oxygen saturation at margin of active duodenal ulcers may lead to slow healing.

This study tested the hypothesis that reduced perfusion of a duodenal ulcer margin (ie, the mucosa 1–2 mm from the edge of the ulcer base) is associated with slow healing. Reflectance spectrophotometric measurement of indices of mucosal hemoglobin concentration (IHB) and mucosal hemoglobin oxygen saturation (ISO2) were obtained endoscopically in 21 patients at the ulcer margin and the adjacent mucosa (ie, the mucosa 1–2 cm from the edge of the ulcer base). In 17 patients with adequate follow-up, stepwise multilinear regression analysis revealed a significantly negative correlation (r=s-0.69, P < 0.05) between ISO2 at the ulcer margin minus ISO2 at the adjacent mucosa (δISO)2 and ulcer healing time. In addition, smoking, being black, and early relapse since the last ulcer attack were found to be associated with increased duration required for healing. The results of this pilot study suggest factors, in addition to smoking, that may have to be considered in future studies concerned with duodenal ulcer healing.