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Cholinergic, gastrinergic, histaminergic and metabolic pathways of PGI2-induced gastric cytoprotection in ethanol-induced mucosal damage in rats
April 1995
Esophageal, Gastric, and Duodenal Disorders
• GLOBUS SENSATION - AN INDICATOR OF PHARYNGOESOPHAGEAL DYSFUNCTION. G MnseL Th-A Abatzi, T Wenzel, U Weber, C Schneider, P Pokieser, W Schima, D-M Denk, M Kutilek, HBergmann, G Stacher. Psychophysiology Unit and Depts. of Gastroenterology, Radiology, Psychiatry, Otolaryngulogy, and Biomedical Physics, University of Vienna, Vienna, Austria. The globus sensation is often viewed as pointing at a conversion disorder or other psychiatric disturbance. We studied 67 females and 21 males (age 22-70 yr, median 42 yr) presenting with the sensation as primary symptom. All underwent a thorough history taking as well as otolaryngological, videocinematographic, and manometric examinations of pharynx and esophagus When indicated, 24-hour pH-metry, scintigraphic quantifications of esophageal bolus transport and gastric emptying, esophagogastroscopy, and other examinations were performed. The investigations revealed that 3 patients had glottal and 1 velopalatinal insufficiency, 1 laryngeal penetration of ingesta, 4 only partial opening of the pharyngoesophageal sphincter upon swallowing, 3 dental malocclusion, 6 sinusitis, 2 laryngeal edema, 2 desiccated pharyngeal mucosa, 10 pharyngoceles, 1 pharyngeal polyps, 17 pharyngitis 5 chronic tonsillitis, 10 hyperplastic lingual tonsils, 11 cervical spondylitis or osteochondritis, 3 cervical spondylophytes, and 21 diffuse struma or thyroidal adenomata. Regarding esophagus and stomach, 24 patients had achalasia I diffuse esophageal spasms, 3 nutcracker esophagus, 32 nonspecific motor abnonnalmes, 5 adynamlc esophagus, ~1 abnormal gastroesophageal reflux (6 also esophagitis), 2 stenoses at the esophagogastric junction, 9 hiatal hernias, 3 massively delayed gastric emptying, and 12 antral gastritis. Of those having esophageal motor abnormalities, 2 also had a Zenker diverticulum, 7 pharyngoceles, 3. glottal insufficiency 2 incomplete pharynguesophageal sphincter opemng, 2 malocclusion, 3 sinusitis, 3 tonsillitis, 12 pharyngitis, 3 cervical spondylosis, 16 goiter or thyroida! adenomata, 1 distal esophageal stenosis, 9 hiatal hernia 2 delayed gastric emptying and 9 gastritis. Psychometric investigations in 82 patients showed no higher scores for depression (Beck), state and trait anxiety (Spielberger), as well as hysteria and hypochondriasis (Minnesota Multiphasic Personality Inventory) than in 83 patients with other gastrointestinal disorders. Psychiatric interviews with 63 patients showed that 36 met DSM-III-R criteria for psychiatric disorders. However, neither this nor the psYchometric findings bore any relationship to the intensity or frequency of occurrence of the globus sensation. The results suggest that the globus sensation has to be viewed as an indicator of oropharyngeal abnormalities as well as disordered pharyngeal, esophageal and possibly gastric motor function. •
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CHOLINERGIC, GASTRINERGIC, lllSTAMINERG1C AND METABOLIC PATIIWAYS OF PGIz-INDUCEI) GASTRIC CYTOPROTECTION 1N ETIIANOL-1NDUCED MUCOSAL DAMAGE IN RATS. Gv. M6zsik, G. Slit/l, A. Kirfiy, A. Vincze, OME. AbdeI-Salam. O. Karfidi. 1st Dept. of Medicine, Univ. Med. Sch., P6cs, thmgary. The aim of the present stnd~ is to investigate the nature of PGI 2induced gastric cytoprolection on the rat gastric mucosal damage in a non-acid-dependent-experimental model. The gastric mncosal damage was produced by intragastric administratinn of 1 ml 96% ETOII. The animals were sacrificed at I hoar after ETOII administration~ when the gastric mucosal damage was measured. The pentagaslrin (Pel>tavhm, ICi. England), histamine (Peremin, Chinnin, llungary), 2,4-dinitraphenol (Ilopkin,s and Williams Ltd, USA) were given snbcntanenusly at 30 rain before administration of ETOII without and with PGI 2 (51ag/kg sc.=EDs0) (Chinoin, Bndapest, lhmgary). The affinity and intrinsic activity curves were calcnlated according to Cs,'iky (1969). The intrinsic activit~ (~+) of pentagastrin was taken as 1.00 on stimulation of PGI2-indnced eytoprotection. The values of p132 (the dose necessary to produce 50% stimulation in the affinity curves) and pA2 I the dose necessary to prnduee 50% stimnlation of the intrinisic activity cnrvesl were calculated from the affinity and intrinsic activity curves. Results:l I The Pt;iz-indnced gastric cytoproteetion was only obt~rined in rats with intact wtgns; 21 The order of stimalatory componnds (based nn wdnes of pl)2, ~. pA2 for drugs stimulating PGI2-indnced gastric cytoproteclion) is presented below: Affinity Intrinsic activity pD 2 oL~ pA2 A B A B A B llistamine 4.70 4.70 0.40 0.05 3.75 3.75 Dinitrophenol 5.10 5.00 0.78 0.82 5.12 4.95 Pentagastrin 7.12 7.12 1.00 1.00 7.00 7.25 A: number of gastric lesions. B: severity of gastric lesions. Conclnsions: 1)Intact vaglls is necessary to obtain gastric cytoprnteetion by PGI2 in ETOtl-model; 2)Pentagastrin and histamine stimulates the PGI 2 gastric cytoprotection; 3) Stinmlatory efi¥ct of dinitrnphennl (by inhibiting cell respiration) appears as direct metabolic action. The study was supported by lhmgarian Natinnal Research Fund '~O'I'KA N~ 2466) and Ministr~ of ltealth and Welfare (ETT-03 660/93~.
A171
HELICOBACTER PYLORI INFECTION INDUCES GASTRIC EPITHELIAL APOPTOSIS IN VIVO SF Moss, J Calam, B Agarwal, S Wang, PR Holt. Dept of Medicine, St.Luke's-Roosevelt Hospital Center/Columbia University, NY & Hammersmith Hospital, London, UK.
H pylori gastritis is an early stage in the development of most gastric cancers but how H pylon contributes to carcinogenesis is unclear. One possible contributory factor is epithelial hyperproliferation, which may itself be a response to increased epithelial programmed cell death (apoptosis) induced by H pyloN. To investigate this hypothesis, we evaluated the degree of epithelial apoptosis in 16 patients with Hpylori antral gastritis, before and after triple therapy. Eradication was confirmed if the culture, urease test and histology were all negative 1 month after therapy, Cells undergoing apoptosis were identified by insitu end-labelling with terminal deoxynucleotidyl ¢ransferase and digoxigenin-dUTP on 4#m sections of formalin-fixed antral biopsies. At least 300 epithelial cells were counted and the number of positive cells per 100 cells was expressed as the apoptotic index. Histological gastritis in the H/E section was scored from 0-10 (Rauws et al. Gastroenterology 1988;94:33). Results: Before treatment apoptotic cells were identified in the surface epithelium and throughout the entire gland. However, following t h e successful eradication of H pylori, in 12 patients, apoptotic cells were only seen rarely, in all cases located superficially. In the patients in whom H pylori was successfully eradicated there was a fall in the median apoptotic epithelial cell index from 7.5% (range 0-44) pretreatment to 1.5% (0-18), p < 0.02 and a fall in median gastritis score from 3.5 (1-8) to 1 (1-6), p < 0.05. Overall, there was no correlation between the apoptotic index and the total gastritis score nor with the neutrophil score, implying that inflammation was not directly responsible for the increased apoptosis. Conclusion: These results show that the eradication of Hpylori reduces the epithelial apoptotic index and imply that H pylori induces gastric epithelial apoptosis. This may be important in the induction of neoplasia by chronic H pylori infection.
BOMBESIN/GRP DOES NOT ALTER ACID SECRETION IN I S O L A T E D M O U S E GASTRIC GLANDS. MJ Muller, I Padol, RH Hunt., IDRP M e M a s t e r University Medical Centre, Hamilton, Ontario, Canada.
Bombesin, also known as gastrin releasing peptide (GRP), has recently b e e n used to document an increase in gastric acid secretion between Helicobacter pylor/-positive duodenal ulcer patients and H.pylorinegative control individuals. The effects of this neuropeptide on gastric acid secretion are complex, involving actions in both the antrum and fandus. In the mouse fundus, bombesin/GRP inhibits basal and histamine-stimulated acid secretion by releasing somatostatin and possibly by acting directly on the parietal cell (AM.J.Physiol.260,G156-G160,1991). The aim of the present study was to investigate the direct effect of b o m b e s i n / G R P on acid secretion in an isolated gastric gland model in the mouse. Methods: The isolated mouse gastric gland preparation was obtained by collagenaso digestion of the mucosa of the mouse stomach. The preparation consisted of gastric glands (approximate 90% total cell number) and a few free cells. The effect of bombesin/GRP and somatostatin on histamine stimulated acid secretion, measured indirectly by the accumulation of the weak base 14C~aminopyrine, was studied by pre-incubating the preparation with the peptide for 30 min. Results: Pre-incubating isolated mouse gastric glands for 30 rain with a large range of concentrations of bombesin/GRP (0.1 nM to I ~M) did not increase or decrease basal acid secretion or acid secretion stimulated with a maximally effective concentration of histamine (100 {*M). On the other hand, somatostatin-14 was a potent inhibitor of histamine (100 #M), stimulated acid secretion in this preparation, confirming our previous findings. Therefore, it is unlikely that the lack of effect of bombesin/GRP is due to selective destruction of inhibitory peptide receptors in mouse parietal cells. Conclusion: Bombesin/GRP-mediated inhibition of basal and histaminestimulated gastric acid secretion in the mouse does not involve a direct effect of this neuropeptide on the parietal cell.
Esophageal, Gastric, and Duodenal Disorders
• GLOBUS SENSATION - AN INDICATOR OF PHARYNGOESOPHAGEAL DYSFUNCTION. G MnseL Th-A Abatzi, T Wenzel, U Weber, C Schneider, P Pokieser, W Schima, D-M Denk, M Kutilek, HBergmann, G Stacher. Psychophysiology Unit and Depts. of Gastroenterology, Radiology, Psychiatry, Otolaryngulogy, and Biomedical Physics, University of Vienna, Vienna, Austria. The globus sensation is often viewed as pointing at a conversion disorder or other psychiatric disturbance. We studied 67 females and 21 males (age 22-70 yr, median 42 yr) presenting with the sensation as primary symptom. All underwent a thorough history taking as well as otolaryngological, videocinematographic, and manometric examinations of pharynx and esophagus When indicated, 24-hour pH-metry, scintigraphic quantifications of esophageal bolus transport and gastric emptying, esophagogastroscopy, and other examinations were performed. The investigations revealed that 3 patients had glottal and 1 velopalatinal insufficiency, 1 laryngeal penetration of ingesta, 4 only partial opening of the pharyngoesophageal sphincter upon swallowing, 3 dental malocclusion, 6 sinusitis, 2 laryngeal edema, 2 desiccated pharyngeal mucosa, 10 pharyngoceles, 1 pharyngeal polyps, 17 pharyngitis 5 chronic tonsillitis, 10 hyperplastic lingual tonsils, 11 cervical spondylitis or osteochondritis, 3 cervical spondylophytes, and 21 diffuse struma or thyroidal adenomata. Regarding esophagus and stomach, 24 patients had achalasia I diffuse esophageal spasms, 3 nutcracker esophagus, 32 nonspecific motor abnonnalmes, 5 adynamlc esophagus, ~1 abnormal gastroesophageal reflux (6 also esophagitis), 2 stenoses at the esophagogastric junction, 9 hiatal hernias, 3 massively delayed gastric emptying, and 12 antral gastritis. Of those having esophageal motor abnormalities, 2 also had a Zenker diverticulum, 7 pharyngoceles, 3. glottal insufficiency 2 incomplete pharynguesophageal sphincter opemng, 2 malocclusion, 3 sinusitis, 3 tonsillitis, 12 pharyngitis, 3 cervical spondylosis, 16 goiter or thyroida! adenomata, 1 distal esophageal stenosis, 9 hiatal hernia 2 delayed gastric emptying and 9 gastritis. Psychometric investigations in 82 patients showed no higher scores for depression (Beck), state and trait anxiety (Spielberger), as well as hysteria and hypochondriasis (Minnesota Multiphasic Personality Inventory) than in 83 patients with other gastrointestinal disorders. Psychiatric interviews with 63 patients showed that 36 met DSM-III-R criteria for psychiatric disorders. However, neither this nor the psYchometric findings bore any relationship to the intensity or frequency of occurrence of the globus sensation. The results suggest that the globus sensation has to be viewed as an indicator of oropharyngeal abnormalities as well as disordered pharyngeal, esophageal and possibly gastric motor function. •
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.
•
'
1
CHOLINERGIC, GASTRINERGIC, lllSTAMINERG1C AND METABOLIC PATIIWAYS OF PGIz-INDUCEI) GASTRIC CYTOPROTECTION 1N ETIIANOL-1NDUCED MUCOSAL DAMAGE IN RATS. Gv. M6zsik, G. Slit/l, A. Kirfiy, A. Vincze, OME. AbdeI-Salam. O. Karfidi. 1st Dept. of Medicine, Univ. Med. Sch., P6cs, thmgary. The aim of the present stnd~ is to investigate the nature of PGI 2induced gastric cytoprolection on the rat gastric mucosal damage in a non-acid-dependent-experimental model. The gastric mncosal damage was produced by intragastric administratinn of 1 ml 96% ETOII. The animals were sacrificed at I hoar after ETOII administration~ when the gastric mucosal damage was measured. The pentagaslrin (Pel>tavhm, ICi. England), histamine (Peremin, Chinnin, llungary), 2,4-dinitraphenol (Ilopkin,s and Williams Ltd, USA) were given snbcntanenusly at 30 rain before administration of ETOII without and with PGI 2 (51ag/kg sc.=EDs0) (Chinoin, Bndapest, lhmgary). The affinity and intrinsic activity curves were calcnlated according to Cs,'iky (1969). The intrinsic activit~ (~+) of pentagastrin was taken as 1.00 on stimulation of PGI2-indnced eytoprotection. The values of p132 (the dose necessary to produce 50% stimulation in the affinity curves) and pA2 I the dose necessary to prnduee 50% stimnlation of the intrinisic activity cnrvesl were calculated from the affinity and intrinsic activity curves. Results:l I The Pt;iz-indnced gastric cytoproteetion was only obt~rined in rats with intact wtgns; 21 The order of stimalatory componnds (based nn wdnes of pl)2, ~. pA2 for drugs stimulating PGI2-indnced gastric cytoproteclion) is presented below: Affinity Intrinsic activity pD 2 oL~ pA2 A B A B A B llistamine 4.70 4.70 0.40 0.05 3.75 3.75 Dinitrophenol 5.10 5.00 0.78 0.82 5.12 4.95 Pentagastrin 7.12 7.12 1.00 1.00 7.00 7.25 A: number of gastric lesions. B: severity of gastric lesions. Conclnsions: 1)Intact vaglls is necessary to obtain gastric cytoprnteetion by PGI2 in ETOtl-model; 2)Pentagastrin and histamine stimulates the PGI 2 gastric cytoprotection; 3) Stinmlatory efi¥ct of dinitrnphennl (by inhibiting cell respiration) appears as direct metabolic action. The study was supported by lhmgarian Natinnal Research Fund '~O'I'KA N~ 2466) and Ministr~ of ltealth and Welfare (ETT-03 660/93~.
A171
HELICOBACTER PYLORI INFECTION INDUCES GASTRIC EPITHELIAL APOPTOSIS IN VIVO SF Moss, J Calam, B Agarwal, S Wang, PR Holt. Dept of Medicine, St.Luke's-Roosevelt Hospital Center/Columbia University, NY & Hammersmith Hospital, London, UK.
H pylori gastritis is an early stage in the development of most gastric cancers but how H pylon contributes to carcinogenesis is unclear. One possible contributory factor is epithelial hyperproliferation, which may itself be a response to increased epithelial programmed cell death (apoptosis) induced by H pyloN. To investigate this hypothesis, we evaluated the degree of epithelial apoptosis in 16 patients with Hpylori antral gastritis, before and after triple therapy. Eradication was confirmed if the culture, urease test and histology were all negative 1 month after therapy, Cells undergoing apoptosis were identified by insitu end-labelling with terminal deoxynucleotidyl ¢ransferase and digoxigenin-dUTP on 4#m sections of formalin-fixed antral biopsies. At least 300 epithelial cells were counted and the number of positive cells per 100 cells was expressed as the apoptotic index. Histological gastritis in the H/E section was scored from 0-10 (Rauws et al. Gastroenterology 1988;94:33). Results: Before treatment apoptotic cells were identified in the surface epithelium and throughout the entire gland. However, following t h e successful eradication of H pylori, in 12 patients, apoptotic cells were only seen rarely, in all cases located superficially. In the patients in whom H pylori was successfully eradicated there was a fall in the median apoptotic epithelial cell index from 7.5% (range 0-44) pretreatment to 1.5% (0-18), p < 0.02 and a fall in median gastritis score from 3.5 (1-8) to 1 (1-6), p < 0.05. Overall, there was no correlation between the apoptotic index and the total gastritis score nor with the neutrophil score, implying that inflammation was not directly responsible for the increased apoptosis. Conclusion: These results show that the eradication of Hpylori reduces the epithelial apoptotic index and imply that H pylori induces gastric epithelial apoptosis. This may be important in the induction of neoplasia by chronic H pylori infection.
BOMBESIN/GRP DOES NOT ALTER ACID SECRETION IN I S O L A T E D M O U S E GASTRIC GLANDS. MJ Muller, I Padol, RH Hunt., IDRP M e M a s t e r University Medical Centre, Hamilton, Ontario, Canada.
Bombesin, also known as gastrin releasing peptide (GRP), has recently b e e n used to document an increase in gastric acid secretion between Helicobacter pylor/-positive duodenal ulcer patients and H.pylorinegative control individuals. The effects of this neuropeptide on gastric acid secretion are complex, involving actions in both the antrum and fandus. In the mouse fundus, bombesin/GRP inhibits basal and histamine-stimulated acid secretion by releasing somatostatin and possibly by acting directly on the parietal cell (AM.J.Physiol.260,G156-G160,1991). The aim of the present study was to investigate the direct effect of b o m b e s i n / G R P on acid secretion in an isolated gastric gland model in the mouse. Methods: The isolated mouse gastric gland preparation was obtained by collagenaso digestion of the mucosa of the mouse stomach. The preparation consisted of gastric glands (approximate 90% total cell number) and a few free cells. The effect of bombesin/GRP and somatostatin on histamine stimulated acid secretion, measured indirectly by the accumulation of the weak base 14C~aminopyrine, was studied by pre-incubating the preparation with the peptide for 30 min. Results: Pre-incubating isolated mouse gastric glands for 30 rain with a large range of concentrations of bombesin/GRP (0.1 nM to I ~M) did not increase or decrease basal acid secretion or acid secretion stimulated with a maximally effective concentration of histamine (100 {*M). On the other hand, somatostatin-14 was a potent inhibitor of histamine (100 #M), stimulated acid secretion in this preparation, confirming our previous findings. Therefore, it is unlikely that the lack of effect of bombesin/GRP is due to selective destruction of inhibitory peptide receptors in mouse parietal cells. Conclusion: Bombesin/GRP-mediated inhibition of basal and histaminestimulated gastric acid secretion in the mouse does not involve a direct effect of this neuropeptide on the parietal cell.