Heparin inhibits protective effect of ischemic preconditioning in ischemia/reperfusion-induced acute pancreatitis.

UNLABELLED: Previous studies have shown that pancreatic ischemic preconditioning or heparin, applied before induction of acute pancreatitis inhibit the development of this disease and accelerate pancreatic recovery. The aim of the study was to determine the influence of treatment with heparin on protective effect of ischemic preconditioning (IP) in ischemia/reperfusion-induced acute pancreatitis. Heparin was administered twice, before and during induction of acute pancreatitis. IP was performed by short-term clamping of celiac artery, 30 min before induction of acute pancreatitis. Acute pancreatitis was induced in rats by clamping of inferior splenic artery for 30 min followed by reperfusion. Rats were sacrificed after 6-h and 24-h reperfusion. RESULTS: IP applied alone caused a mild pancreatic damage associated with a limited increase in plasma amylase activity, concentration of pro-inflammatory interleukin-1ß and plasma level of D-dimer. Pretreatment with heparin or IP applied alone reduced the severity of acute pancreatitis. Both these procedures caused a similar reduction in plasma lipase, amylase and interleukin-1ß, as well as in histological signs of pancreatic damage. These changes were associated with partial reversion of the pancreatitis-evoked fall of pancreatic blood flow and DNA synthesis. Combination of heparin plus IP reduced the protective effect of heparin or IP applied alone. It was manifested by an increase in pancreatic damage and plasma level of lipase, amylase and interleukin-1ß, as well as by reduction in pancreatic DNA synthesis and plasma concentration of D-dimer and interleukin-10. CONCLUSIONS: heparin abolishes the protective effect of ischemic preconditioning in ischemia reperfusion-induced pancreatitis. This observation suggests that initial clot formation is necessary to induce pancreatic protection by IP.

Pretreatment with obestatin inhibits the development of cerulein-induced pancreatitis.

Obestatin is a peptide derived from the proghrelin, a common prohormone for ghrelin and obestatin. Obestatin, like the ghrelin has been originally extracted from rat stomach, and the stomach seems to be a major source of circulating obestatin. Previous studies have shown that administration of ghrelin exhibits protective effect in the pancreas, inhibiting the development of acute pancreatitis. Recent study has shown that obestatin promotes survival of beta-cells and pancreatic islets. Aim of the present study was to investigate the influence of obestatin administration on the development of cerulein-induced pancreatitis. Studies were performed on male Wistar rats. Acute pancreatitis was induced by cerulein given intraperitoneally 5 times at a dose of 50 microg/kg/dose with 1-h intervals. Obestatin was injected twice intraperitoneally at the dose of 4, 8 or 16 nmol/kg/dose. In control saline-treated rats, obestatin was without effect on pancreatic morphology, serum activity of pancreatic enzymes, serum level of pro-inflammatory interleukin-1beta or pancreatic cells proliferation. In animals with induction of acute pancreatitis, morphological examination showed that administration of obestatin decreased pancreatic leukocyte infiltration and vacuolization of acinar cells. These effects were accompanied by reduction in the pancreatitis-evoked increase in serum level of pancreatic digestive enzymes, lipase amylase and poly-C ribonuclease. Obestatin administered at the highest dose of 16 nmo/kg/dose reduced serum activity of these enzymes by 33, 42 and 44%, respectively. Also serum concentration of pro-inflammatory interleukin-1beta was decreased by obestatin in rats with acute pancreatitis; whereas the pancreatitis-evoked decrease in pancreatic blood flow and pancreatic DNA synthesis was partially reversed. Administration of obestatin reduces the severity of cerulein-induced acute pancreatitis. This effect is related, at least in part, to the improvement of pancreatic blood flow and reduction in proinflammatory interleukin-1beta release.

Treatment with ghrelin accelerates the healing of acetic acid-induced gastric and duodenal ulcers in rats.

UNLABELLED: Recent studies have shown that ghrelin exhibits gastroprotective effects. The aim of present study was to examine the influence of ghrelin administration on the healing of chronic gastric and duodenal ulcers and to evaluate the role of growth hormone (GH) and insulin-like growth factor-1 (IGF-1) in this process. In pituitary-intact or hypophysectomized rats, chronic gastric and duodenal ulcers were induced by acetic acid. After induction of ulcers, rats were treated intraperitoneally twice a day with saline, ghrelin (4, 8 or 16 nmol/kg/dose) or IGF-1 (20 nmol/kg/dose) for six or ten days. In animals with intact pituitary, treatment with ghrelin increased serum level of GH and IGF-1. These effects were accompanied by the increase in mucosal cell proliferation, mucosal blood flow and healing rate of gastric and duodenal ulcers. After hypophysectomy, the significant increase in serum level of endogenous ghrelin was observed, but the healing of gastric and duodenal ulcers was delayed. This effect was accompanied by a significant decrease in serum concentration of endogenous GH and IGF-1, and reduction in mucosal blood flow and DNA synthesis. In hypophysectomized rats, administration of exogenous ghrelin was without any effect on serum level of GH and IGF-1, healing rate of gastroduodenal ulcers or mucosal cell proliferation. In contrast to this effect, administration of IGF-1 increased mucosal cell proliferation, healing rate of gastroduodenal ulcers and mucosal blood flow in hypophysectomized rats. CONCLUSION: Treatment with ghrelin accelerates healing of chronic gastroduodenal ulcers and this effect is mediated by the release of endogenous GH and IGF-1.

Role of afferent nerves and sensory peptides in the mediation of hepatic artery buffer response.

Intrahepatic arteries are richly innervated by both adrenergic and sensory vanilloid-sensitive (capsaicin-sensitive) fibers. Stimulation of capsaicin sensitive fibers has been shown to dilate the intrahepatic vessels by both releasing sensory neuropeptides and by modulating the adrenergic tone. However the participation of capsaicin-sensitive fibers in the mediation of the hepatic artery buffer response (HABR) has not been investigated yet. To explore the involvement of sensory innervation and sensory neuropeptides in the HABR, the experiments were performed on capsaicin-denervated Wistar rats. In addition, we used selective CGRP and tachykinin receptor antagonists to test the participation of CGRP, substance P and NK-A in HABR in the rat. In anesthetized rats the hepatic artery blood flow (HABF), microcirculatory hepatic blood flow (HBF) and portal blood flow (PBF) were determined. The HABR was induced by partial occlusion of the portal vein and maintaining the PBF at 10% of its control preocclusive value. In the control HABR the hepatic artery blood flow increased by 89% (p< 0.005) whilst the HBF at the same time decreased by 32% (p< 0.005) in comparison to preocclusive HABF and HBF values. In sensory-denervated rats the resting HBF and PBF were increased by 23% (p< 0.05) and 34% (p< 0.05), respectively in comparison to the control HBF and PBF values. In this group the induction of the HABR increased the hepatic artery blood flow by only 55% (p< 0.05), whilst the HBF was reduced by 45% (p< 0.05). Pretreatment with CGRP 8-37 (CGRP receptor antagonist) and NK-1 but not NK-2 nor NK-3 receptor antagonists significantly reduced the HABF by 43% (p< 0.05) and 25% (p< 0.05) as compared to the HABF value in the control HABR group. These findings support the hypothesis that the hepatic artery buffer response induced by reduction of the portal inflow to the liver by 90% is partially mediated by activation of capsaicin-sensitive sensory fibers in the liver, probably due to local tissue ischemia and hypoxia. The observed vasodilation in the vascular bed of the hepatic artery is due to stimulation of CGRP and NK-1 receptors.

Histamine H3 receptors modulate reactive hyperemia in rat gut.

Reactive hyperemia (RH) is an abrupt blood flow increase following release from mechanical occlusion of an artery, with restoration of intra-arterial pressure. The mechanism of this postocclusion increase in blood flow in the gut is multifactorial. Relaxation of intestinal resistance vessels, observed during RH, may involve myogenic, metabolic, hormonal and neurogenic factors. Evidence exists that histamine is an important endogenous mediator of various functions of the gut, including blood flow. The vascular effects of histamine in the intestinal circulation are due its agonistic action on histamine H1, H2 and H3 receptors. In the present study the hypothesis was tested that peripheral histamine H3 receptors are involved in the mediation of RH in the intestinal circulation. In anesthetized rats, anterior mesenteric artery blood flow (MBF) was determined with ultrasonic Doppler flowmeter, and arterial pressure (AP) was determined with a transducer. The increase in the volume of blood accumulating during RH (RH-volume), the peak increase of arterial blood flow (RH-peak response) and the duration of the hyperemia (RH-duration) were used to quantify RH after occluding the anterior mesenteric artery for 30, 60 and 120 s. Hyperemia parameters were determined before and after administration of the selective histamine H3 receptor antagonist clobenpropit. Pretreatment with clobenpropit was without any effect on control MBF and AP but significantly reduced most of RH responses. These findings support the hypothesis that histamine H3 receptors do not play any role in the control of intestinal vasculature at basal conditions but these receptors participate in the intestinal hyperemic reaction in response to complete temporal intestinal ischemia.

Pancreatic damage and regeneration in the course of ischemia-reperfusion induced pancreatitis in rats.

The objective of this study was to assess the biochemical and histological signs of pancreatic damage development and pancreatic recovery in the course of ischemia-reperfusion induced pancreatitis. Acute pancreatitis was induced in rats by limitation of pancreatic blood flow (PBF) in inferior splenic artery for 30 min using microvascular clips, followed by reperfusion. Rats were sacrificed at the time: 1 h, 12 h, 24 h, and 2, 3, 5, 7, 10, 14, 21 and 28 days after ischemia. PBF was measured using laser Doppler flowmeter. Plasma amylase, interleukin 1beta (IL-1beta) and interleukin 10 (IL-10) concentration, pancreatic DNA synthesis, as well as, morphological features of pancreatic damage were examined. Ischemia with reperfusion caused acute necrotizing pancreatitis followed by pancreatic regeneration. After removal of microvascular clips, PBF was reduced and the maximal fall of PBF was observed 24 h after ischemia, then PBF grew reaching the control value at 28th day. Plasma amylase activity was increased between 12th h and 3rd day with maximum at 24 h after ischemia. Also plasma IL-1beta and IL-10 were elevated with maximal value at the first and second day after ischemia, respectively. DNA synthesis was maximally reduced at the first day (by 70%) and from second day the reversion of this tendency was observed with full restoration of pancreatic DNA synthesis within four weeks. Morphological features of pancreatic tissue showed necrosis, strongly pronounced edema and leukocyte infiltration. Maximal intensity of morphological signs of pancreatic damage was observed between first and second day of reperfusion. During pancreatic regeneration between second and tenth day after ischemia the temporary appearance of chronic pancreatitis-like features such as fibrosis, acinar cell loss, formation of tubular complexes and dilatation of ducts was observed. The regeneration was completed within four weeks after pancreatitis development. We conclude that partial and temporary pancreatic ischemia followed by reperfusion causes acute necrotizing pancreatitis with subsequent regeneration within four weeks. Pancreatic repair after necrotizing pancreatitis is connected with the increase in plasma IL-10 concentration and transitory formation of tubular complexes.

Involvement of cyclooxygenase-derived prostaglandin E2 and nitric oxide in the protection of rat pancreas afforded by low dose of lipopolysaccharide.

Prostaglandins (PG), the products of arachidonate metabolism through cyclooxygenase (COX) pathway, protect the pancreas from the acute damage. The existence of two isoforms of COX was documented including: COX-1, present in normal tissues and COX-2, expressed at the site of inflammation, such as induced by bacterial lipopolysaccharide (LPS). Pretreatment with low dose of LPS and activation of nitric oxide (NO) synthase (NOS) has been shown to prevent the injury caused by caerulein-induced pancreatitis (CIP) in the rat. The aim of this study was to investigate the role of COX-1 and COX-2 in the LPS-induced protection of the pancreas against CIP and the involvement of NOS in the activation of COX-PG system in the rats with CIP. CIP was produced by subcutaneous (s.c.) infusion of caerulein (5 microg/kg-h for 5 h) to the conscious rats. Protective dose of LPS, from Escherichia coli, (1 mg/kg) was given intraperitoneally (i.p.) 15 min prior to the start of CIP. Nonselective inhibitor of COX; indomethacin (5 or 10 mg/kg), selective inhibitor of COX-1: resveratrol, or a highly selective inhibitors of COX-2: rofecoxib or NS-398 (2 or 10 mg/kg) were injected i.p. 15 min prior to the administration of LPS. COX-1 or COX-2 mRNA was determined by reverse transcription-polimerase chain reaction (RT-PCR) in the pancreatic tissue. Pancreatic blood flow (PBF) was measured by a laser Doppler flowmetry. PGE2 content in the pancreas was measured by radioimmunoassay. CIP was manifested by an increase of pancreatic weight and plasma amylase activity (by 500% and 700%, respectively) and it was confirmed by histological examination. CIP slightly increased pancreatic PGE2 generation (by 12%) and diminished PBF (by about 40%). LPS (1 mg/kg i.p.), given prior to the start of CIP, increased PGE2 generation in the pancreas (by 45%), reversed the histological manifestations of pancreatitis, reduced the rise in amylase blood level and improved PBF. Administration of nonselective inhibitor of COX; indomethacin (5 or 10 mg/kg i.p.) prior to the injection of LPS abolished its protective effects on CIP and reduced pancreatic PGE2 generation. Selective inhibitor of COX-1; resveratrol (10 mg/kg i.p.) given prior to the injection of LPS reversed its protective effects against CIP. Pretreatment with a selective inhibitors of COX-2: rofecoxib or NS-398 (10 mg/kg) attenuated LPS-induced pancreatic protection in the CIP rats. COX-1 expression was detected in the intact pancreas and was not significantly changed by CIP, LPS, indomethacin, NS-389 and their combination, while COX-2 mRNA expression appeared in the pancreas of ratssubjected to CIP and was significantly increased after LPS injection to these rats. Addition of selective COX-2 inhibitor; NS-389, or nonselective inhibitor of COX; indomethacin, enhanced COX-2 mRNA expression in the rats with CIP pretreated with LPS. Pretreatment of the rats with inhibitor of NOS; L-NNA (20 mg/kg i.p.), given together with LPS, 15 min prior to the start of caerulein overstimulation, resulted in complete reversion of LPS-induced pancreatic protection and decreased PGE2 generation stimulated by LPS. Addition to L-NNA of the substrate for NOS; L-arginine (100 mg/kg i.p.), restored pancreatic protection afforded by low dose of LPS and increased pancreatic PGE2 level in the rats with CIP. We conclude that: 1. increased pancreatic PGE2 generation, induced by low dose LPS pretreatment, contributes to the pancreatic resistance to acute damage produced by caerulein overstimulation and 2. the NO-system is involved in above stimulation of PGE2 generation and pancreatic protection against acute damage.

The effect of insulin and sulodexide (Vessel Due F) on diabetic foot syndrome: pilot study in elderly patients.

OBJECTIVE: To assess the efficacy of insulin plus sulodexide (a mixture of 80% heparin-like substances and 20% dermatan sulphate) on diabetic ulcers, and its influence on foot skin microcirculation and diabetic neuropathy. RESEARCH DESIGN AND METHODS: Two groups of diabetic patients, suffering from severe neuropathy and ulceration, were randomly assigned to insulin (I) plus sulodexide (S) (n=12) or insulin plus placebo (P) (n=6) therapy, for 10 weeks. Laser Doppler assessment of foot skin flow (LDF), at rest and 30 or 60 s after arterial occlusion, and nerve conduction tests (sensorial evoked and motoric conduction potentials) have been evaluated in both groups. RESULTS: Postischaemic flow was 2.5 times shorter in ulcerated vs. non-ulcerated feet in diabetic patients. A significant increase in flows after 30 and 60 s ischaemia was detected in both groups at the end of therapy (IS group, ulcerated foot, LDF=60 s: from 99.1+/-14.3 to 218.6+/-28.6 PU, P<.001. IP group=from 110.5+/-13.0 to 164.8+/-15.4 PU, P<.05). The length of reactive hyperaemia was higher in IS vs. IP group (IS: from 30.3+/-2.9 to 43.9+/-2.2 s, P<.001; IP: from 28.7+/-3.0 to 33.3+/-3.3 s, ns). Ninety-two percent of ulcers heals in a mean time of 46.4 days (IS group) vs. 83% and 63.0 days, respectively, in IP group. Nerve conduction studies have not demonstrated within- and between-group differences. CONCLUSIONS: Sulodexide and insulin improve the postischaemic skin flow in ulcerated feet, without affecting nerve conduction tests. The effect of sulodexide results additive to insulin; it is clinically relevant, in the view of the possibility of reducing the time needed to completely heal ulcers. The ultimate validation of these preliminary results requires extensive trials.

Protective action of lipopolysaccharidesin rat caerulein-induced pancreatitis: role of nitric oxide.

Lipopolysaccharides (LPS), the component of the cell wall of gram-negative bacteria, have been implicated in the pathogenesis of acute pancreatitis, but the mechanism of their action on the pancreas has not been fully explored. The aim of this study was to investigate the effects of various doses of LPS on the integrity of intact pancreas and that involved in acute caerulein-induced pancreatitis (CIP) in the rat and to compare these effects with those of nitric oxide (NO) donor, S-nitrose-acetylpenicillamine (SNAP). The expression of constitutive NO synthase (cNOS) and inducible NO synthase (iNOS) mRNA was also examined in the isolated pancreatic acini obtained from the inflamed pancreas of rats treated with LPS. CIP was produced by subcutaneous (s.c.) infusion of caerulein (5 microg/kg.h for 5 h) to conscious rats. Bolus injections of various doses of LPS (0.1, 1, 10, 20 or 40 mg/kg) or SNAP (1.5, 3 or 6 mg/kg) were made intraperitoneally (i.p.) either alone or 30 min prior to s.c. infusion of caerulein to induce CIP. Infusion of caerulein produced acute pancreatitis confirmed by histological examination and manifested by an increase of pancreatic mass (by about 200%). Blood levels of amylase and lipase were augmented by 400 and 800% respectively, whereas the pancreatic blood flow (PBF) was decreased by 50% in rats with CIP. Injection of low doses of LPS (0.1-1 mg/kg i.p.) or SNAP (1.5-3 mg/kg i.p.) 30 min prior to caerulein infusion reversed the harmful effects of pancreatic overstimulation with caerulein and reduced significantly the histological manifestations of CIP such as edema, neutrophil infiltration and vacuolization of the acinar cells. These protective effects of low doses of LPS pretreatment on the pancreas were completely antagonized by the suppression of the activity of NO synthase (NOS) with N(G)-nitro-L-arginine (L-NNA) applied (20 mg/kg i.p.) 15 min prior to the LPS injection. Combination of L-arginine (100 mg/kg i.p.), a substrate for NOS, with L-NNA given prior to low doses of LPS, restored the LPS-induced protection of the pancreas in rats with CIP. In contrast, higher doses of LPS (20-40 mg/kg i.p.) or SNAP (6 mg/kg i.p.), which produced a significant fall of the PBF, did not protect the pancreas against CIP. Administration of various doses of LPS to rats with CIP resulted in significant and dose-dependent stimulation of NO biosynthesis in the isolated acini obtained from the pancreas of these animals. LPS enhanced the expression of both cNOS and iNOS in the pancreatic acini obtained from rats subjected to CIP. The signal for cNOS mRNA was detected in all samples, reaching peak at the protective dose of LPS (1 mg/kg i. p.), while iNOS was overexpressed only at the highest doses of LPS that failed to exhibit the protective activity. We conclude that the pretreatment with low doses of LPS protects the pancreas against the damage provoked by CIP and this effect could be attributed, at least in part, to the activation of L-arginine-NO system in the pancreas.

Role of calcitonin gene related peptide in the modulation of intestinal circulatory, metabolic, and myoelectric activity during ischemia/reperfusion.

Calcitonin gene related peptide (CGRP) is widely distributed in the myenteric neurons along GI tract. Biological effects of CGRP on gastrointestinal tract include increase in the intestinal blood flow, relaxation of the smoth muscle and slight increase in the slow wave amplitude (SWA) with no effect on frequency (SWF) of intestinal myoelectric activity (IMA). The aim of this study was to evaluate the possible protective effects of endo- and exogenous CGRP against ischemia/reperfusion (I/R) induced bowel injury in rats. Experiments were performed on 30 fasted anesthetized Wistar rats. Systemic arterial blood pressure (AP), mesenteric blood flow (MBF) and microcirculatory intestinal blood flow (LDBF) were measured. Oxygen consumption (VO2) was estimated from MBF and mesenteric AVO2 difference. IMA was recorded by 4 monopolar electrodes in proximal jejunum and analyzed using computer program with Fourrier analysis of SWF. Control ischemia induced by 30 min total occlusion of anterior mesenteric artery (AMA) reduced SWA by 25+/-5% and SWF by 24+/-4%. At the end of 60 minute reperfusion period SWA and SWF values were restored to control values but SWF showed instability. At the 15th minute of reperfusion period MBF, LDBF and VO2 increased to 109+/-6, 119+/-11 and 120+/-7% of control values respectively. Infusion of exogenous CGRP (0,16 microg/kg/min i.a.) increased MBF, LDBF and VO2 by 26+/-6, 31+/-9 and 20+/-4% respectively in comparison to control values. In the same experimental group SWA increase of 14% was observed with not significant changes in SWF. In the group where CGRP was administrated before and during 30 min period of intestinal ischemia MBF, LDBF and VO2 values at 15th minute of reperfusion were significantly higher by 24+/-6, 32+/-7 and 17+/-5% respectively in comparison to the values observed in the control reperfusion. In that group SWA values were restored to preocclusion values at 15th minute of reperfusion yet and SWF showed much more stability. Infusion of CGRP receptor antagonist (CGRP 8-37) reduced MBF, LDBF and VO2 by 12+/-7, 14+/-8 and 11+/-6% respectively (differences not significant versus control). In I/R group when CGRP 8-37 was given hemodynamic parameters (during reperfusion) were significantly lower and IMA parameters were not restored to preocclusion values. We conclude, that endogenous and exogenous CGRP attenuate circulatory parameters of the small intestine during ischemia and reperfusion. A direct correlation exists between hemodynamic, metabolic and myoelectric effects of CGRP.

Myoelectric bowel activity in ischemia/reperfusion damage. Role of sensory neurons.

The present knowledge indicates that afferent sensory neurons (C-fibres) play an important role in the relationship between intestinal myoelectric activity (IMA) and blood flow (LDBF). The aim of this study was to evaluate the role of C-fibers in myoelectric activity of small intestine during its ischemia and reperfusion. A neurotoxin-capsaicin (CAP) was used to induce functional ablation of afferent sensory neurons. Experiments were performed on 6 groups of anesthetized rats. In the I, II, III group of rats IMA and LDBF were recorded during 100% ischemia induced by AMA 15, 30 and 60 min total occlusion and during 60 min reperfusion period. In group V and VI, IMA and LDBF were registered after intrajejunal placement of 1% CAP. In group IV we measured effects of intraluminal instillation of CAP alone. Intraluminal placement of CAP induced an early increase in slow wave amplitude SWA and slow wave frequency SWF by 35+/-11% and 19+/-10% (p<0.05) with the subsequent decrease in both by 25+/-6 and 24+/-8% (p<0.05) respectively. Short 15 min lasting ischemia induced by 100% occlusion of AMA evoked only a slight increase of SWA. During reperfusion period SWA and SWF returned to the baseline values after 15 min. Total 30 min occlusion decreased SWA and SWF by 25+/-9 and 24+/-6% (p<0.05) respectively. During reperfusion period recovery of IMA parameters to preocclusion values were slower. Intestinal hyperemia was smaller than in previous group. After 60 min lasting intestinal ischemia SWA and SWF were decreased by 58+/-7 and 40+/-6% (p<0.01) respectively. There was no return of IMA parameters to control values. These data demonstrated that intestinal ischemia induces typical changes in the bowel myoelectric activity. These changes possess their own electrical characteristics which can be used in clinical practice for evaluation of the degree ischemically-induced intestinal injury. Capsaicin pretreatment significantly decreased SWA and SWF and LDBF in comparison with those observed in group II and III during 30 and 60 min occlusion and reperfusion period. We conclude that afferent neurons C activated during mesenteric ischemia/reperfusion play an important role in protecting ischemic bowel viability.

[Mechanism of action of neurotensin on microcirculation, metabolism and motility of the small intestine]. / Mechanizm dzialania neurotenzyny na mikrokrazenie, metabolizm oraz motoryke jelita cienkiego.

The aim of this study was to investigate the influence of neurotensin on total microcirculatory blood flow, oxygen consumption, myoelectric activity and motility of the small bowel. The attempts were made to evaluate the role of L-Arginine, NO-system in the mechanism of action of this peptide on microcirculation, metabolism and motor-activity of the intestine. The experiments were performed in two experimental models--dogs and rats. In Vetbutal anesthetised animals the systemic arterial pressure, superior mesenteric artery blood flow, microcirculatory blood flow and myoelectric activity of the small bowel were continuously monitored. In experiments with dogs effective capillary index, arterio-venous oxygen difference and oxygen consumption were analysed. Neurotensin administered intraarterially caused a dose depended increase in total and microcirculatory blood flow in the small bowel in both groups of animals. The experiment with dogs showed the rise of effective capillary index and intestinal oxygen consumption. Administration of neurotensin changed the profile of myoelectric activity of the small bowel similar to that of postprandial hyperaemia. Inhibition of NO-synthase by the intravenous administration of L-NNA reduced significantly the amplitude of neurotensin hyperaemia. Pretreatment with L-Arginine, the substrate of NO-synthase reversed the hemodynamic, but not motility effects of neurotensin. The results presented proved the role of the L-Arginine-NO in circulatory mechanism, but not motility effects of neurotensin.

Nitric oxide as mediator of bradykinin-induced pancreatic circulatory and metabolic responses.

Bradykinin (BK) is an endogenous nonapeptide with potent vasodilator properties of the visceral circulation. BK alters vascular tone via two BK receptor subtypes, B1 and B2. Current experimental evidence suggests that the dilator action of BK in some vessels is mediated primarily by B2 receptor activation. In addition, there are reports that BK increases endothelial generation of vasodilator factors, such as nitric oxide (NO). The present study had two aims. First, to explore the role of BK-receptors in the pancreatic vasodilatatory and metabolic responses to BK. Second aim was to examine whether endogenous NO play a role in the mediation of BK-receptors induced pancreatic circulatory and metabolic activity. In anesthetized dogs, the superior pancreatico-duodenal artery blood flow (SPBF) was measured by ultrasonic blood flowmeter (Transonic System T-206), pancreatic microcirculatory blood flow (PBF) was determined by laser Doppler flowmetry (Periflux 4001 Master). Pancreatic oxygen consumption (PVO2) was calculated as the product of the arteriovenous oxygen difference (AVO2) across the pancreatic circulation and SPBF. Drugs were infused into the superior pancreatico-duodenal artery. BK (0.01-1.0 mg/kg/min) increased maximally SPBF by 180 +/- 15%, PBF by 208 +/- 22% and PVO2 by 145 +/- 11%, respectively. Pretreatment with B2-subtype receptor antagonist, D-Arg, [Hyp3, Thi5,8, D-Phe7] BK inhibited significantly BK-induced increase in SPBF, PBF and PVO2 by 86 +/- 8%, 73 +/- 7% and 85 +/- 6%, respectively. A nitric oxide synthesis inhibitor (L-NNA) administered i.v. at dose of 25 mg/kg 20 min before BK, inhibited significantly the pancreatic hyperemic and metabolic responses. The results presented emphasize an important role of B2 receptors in the mediation of pancreatic circulatory and metabolic responses to bradykinin. Endogenous NO plays a mediatory role in the pancreatic vascular and metabolic responses due to stimulation of B2-receptors.

Role of nitric oxide in the pancreatic vascular and metabolic responses associated with activation of capsaicin-sensitive neurons.

The effects of local periarterial placement of capsaicin upon pancreatic blood flow, oxygen extraction from pancreatic circulation and oxygen consumption by pancreatic tissue were measured in anesthetized dogs. These studies explored also the possible role of endogenous nitric oxide (NO) in the pancreatic vascular and metabolic responses to periarterial capsaicin. In anesthetized dogs, superior pancreatico-doudenal artery blood flow (SPBF) was measured with an ultrasonic blood flowmeter. Microcirculatory pancreatic blood flow (PBF) was measured with laser-Doppler flowmeter. Arteriovenous oxygen difference (AVO2) across the pancreatic circulation was determined spectrophotometrically. Pancreatic oxygen uptake was calculated as the product of AVO2 and SPBF. Capsaicin applied periarterialy induced initial increase in SPBF, PBF and oxygen uptake. The acute capsaicin-induced vascular dilation was followed by steady state response characterized by significant decrease in SPBF, PBF and oxygen uptake. Inhibition of NO synthase by N omega-nitro-L-arginine (L-NNA) induced pancreatic ischemia and hypoxia. After pretreatment with L-NNA the acute capsaicin-induced pancreatic vascular dilation and the increase in pancreatic oxygen uptake were significantly reduced. Above circulatory and metabolic effects of L-NNA were significantly attenuated when administration of L-NNA was combined with L-arginine. The results of these studies indicate that sensory C-fibers at rest and when activated play a role in the control of pancreatic blood flow and tissue oxygenation. These findings support also the hypothesis that NO plays a role in the mediation of pancreatic vasodilatory action of neuropeptides released from sensory C-fibers.

[Vascular factors in the mechanism of acute pancreatitis]. / Czynnik naczyniowy w mechanizmie ostrego zapalenia trzustki.

Clinically, acute pancreatitis can sometimes be seen in local or systemic vascular diseases. Vascular mechanisms of acute pancreatitis are defined as impairment of pancreatic blood inflow and outflow, or disturbance of the pancreatic microcirculation. Pancreatic macro-circulatory flow can be impaired by low flow states such as hypovolemic shock, cardiogenic shock, or by occlusion of pancreatic arteries or veins. The pancreatic microcirculation can be compromised by embolization of microvessels or by induction of endothelial lesions. It is known that pancreatic ischemia is an initiating or promoting factor of acute pancreatitis. It has been proposed that impaired local pancreatic microcirculation results from leukocytes-endothelial cell adhesion and presence of various vasoactive substances or chemical mediators such as histamine, serotonin, prostaglandins, thromboxanes, leukotrienes, platelet-activating factor, endothelin, free radicals, kinins and adhesion glycoproteins.

Role of capsaicin-sensitive neurons in the control of intestinal blood flow and oxygen uptake.

The effects of periarterial placement of capsaicin upon intestinal blood flow, oxygen consumption and distribution of blood flow to the vascular compartments of the gut wall were measured in anesthetized dogs. Total blood flow to the segment of distal ileum was measured with an ultrasonic blood flowmeter and arteriovenous oxygen difference (AVO2) across the intestinal segment was determined spectrophotometrically. Intestinal oxygen uptake was calculated as the product of AVO2 and intestinal blood flow. Intestinal mucosal blood flow was also measured by laser-Doppler flowmeter. Changes in blood flow distribution were estimated from the distribution of radiolabeled microspheres. Capsaicin applied periarterially induced early increase in intestinal blood flow, oxygen uptake and increase in mucosal blood flow which was higher then increase in total blood flow. The acute capsaicin-induced vascular dilation was followed by steady state response characterized by significant decrease in intestinal blood flow, oxygen uptake and redistribution of blood flow away from mucosal-submucosal the intestinal compartment. The results of these studies indicate that sensory C-fibers at rest, and when activated play a role in the control of intestinal blood flow, its distribution among vascular compartments of the intestinal wall and intestinal tissue oxygenation.

Nitric oxide mediates intestinal hyperaemic responses to intraluminal bile-oleate.

It has long been recognized that intestinal blood flow increases at mealtimes. Mesenteric hyperaemia is also evoked by activation of sensory peptidergic nerves. Our studies explored the possible role of endogenous nitric oxide (NO) in the rat intestinal vasodilator response to luminal instillation of an oleic acid plus bile mixture before and after acute intrajejunal instillation of capsaicin and after chronic pretreatment with capsaicin. In anaesthetized rats we measured jejunal blood flow (BF) with an ultrasonic Doppler flowmeter and systemic arterial pressure (AP) with a pressure transducer. Intestinal perfusion with 80 mM oleic acid in bile increased BF by 98 +/- 12%. Instillation of 4 mg of capsaicin into the jejunal lumen initially increased BF by 42 +/- 9% but was followed by vasoconstriction. Inhibition of NO synthase with 25 mg/kg i.v. N-nitro-L-arginine (L-NNA) decreased BF by 27 +/- 5% and increased AP by 37 +/- 11%. After treatment with L-NNA and after acute and chronic administration of capsaicin, the bile-oleate-induced maximal increases in BF above control levels were 42 +/- 7%, 65 +/- 12%, and 58 +/- 8%, respectively. The observed inhibitory effect of L-NNA on the intestinal hyperaemic response to the bile-oleate mixture was reversed by pretreatment with L-arginine (100 mg/kg i.v.). In capsaicin pretreated rats the subsequent bile-oleate-induced hyperaemia was reduced in magnitude but the inhibitory effects of L-NNA were proportionately the same as in animals not receiving capsaicin. These findings support the hypothesis that NO is involved with bile-oleate-induced mesenteric hyperaemia.

Nitric oxide is involved in the mediation of gastric blood flow and tissue oxygenation.

Endogenous nitric oxide which is enzymatically formed by endothelial cells from L-arginine has been implicated in the control of gastrointestinal circulation. Its role in the mediation of gastric tissue oxygenation has not been studied. We investigated the role of NO in the control of gastric blood flow and oxygen uptake. In anesthetized dogs, total gastric blood flow, gastric mucosal blood flow, systemic arterial and portal venous pressures and the arteriovenous oxygen content difference were studied. From these measurements gastric vascular resistance and oxygen consumption were calculated. Administration of NG-nitro-L-arginine (L-NNA) induced gastric tissue ischemia and hypoxia. Both, systemic arterial pressure and gastric vascular resistance were increased. Above hemodynamic and metabolic effects of L-NNA were significantly attenuated when administration of L-NNA was combined with L-arginine. Our findings suggest that endogenous NO is a tonic vasodilator modulating gastric blood flow and oxygen uptake through influence on the gastric microcirculatory structures responsible for vascular resistance and the nutrient circulation.

[Pancreatic blood flow and its regulation]. / Trzustkowy przeplyw krwi i jego regulacja.

Regulatory mechanisms of the pancreatic circulation include general hemodynamics, autonomic nervous system, circulating vasoactive peptides, hormones and certain autoregulatory properties of the pancreatic vasculature. The pancreatic response to sympathetic stimulation is a decrease in pancreatic blood flow (PBF) due to co-release of norepinephrine and NPY. In contrast parasympathetic stimulation increase PBF and this could be attributed to the co-release of acetylcholine, VIP, PHI, GRP and other neuropeptides. Neurotransmitters of the NANC system such as: SP, NK, SK, CCK, VIP, cGRP are also potent neurogenic vasodilatators of the pancreatic vasculature. Control of the pancreatic circulation is related, in part to the exocrine function of the gland. Several factors have been implicated as mediators of functional hyperemia in the pancreas. Among these factors are such low PO2, ions, kinins, PGs, nucleotides and gastrointestinal hormones. Recently endothelium-derived NO has been shown to play an important role in the mediation of the pancreatic vasodilation in response to various endogenous factors.

Microcirculatory and motor effects of endogenous nitric oxide in the rat gut.

The aim of the study was to determine the role of endogenous nitric oxide (NO) as the mediator of intestinal blood flow and motility. Experiments were performed on anesthetized rats. Blood flow in the jejunum was determined by Laser-Doppler flowmeter. Motility was monitored on the basis of changes in intrajejunal pressure. Systemic arterial pressure was also recorded. To investigate the potential role of nitric oxide in the regulation of basal intestinal blood flow and motility the NO synthase inhibitor NG-nitro-L-arginine (L-NNA) was given systemically. Intravenous bolus of L-NNA (15 mg/kg) reduced basal intestinal blood flow and increased both intestinal motility and arterial pressure in the dose-dependent manner. To test the specificity of the NO synthase blockade we administered L-arginine alone or in combination with L-NNA. Pretreatment with L-arginine (100.0 mg/kg i.v.) alone had no major influence but when combined with L-NNA it reversed the intestinal circulatory and motor effects of L-NNA. The results of these studies suggest that endogenous NO exerts a tonic relaxatory influence on the smooth muscle of the intestinal vessels and intestinal wall.