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Prostaglandin E2 inhibits ethanol-induced histamine release from isolated canine gastric mast cells
April 1995
Esophageal, Gastric, and Duodenal Disorders A261
• DECREASE OF INTRAGASTRIC ACIDITY FOLLOWING LOW DOSES OF RANITIDINE JW Wyeth 1, RE_ Pounder 1, CCL Snell2, IRoyal Free Hospital School of Medicine, London, ~;-----~GlaxoResearch & Development Ltd, Grcenford, UK
REGRESSION OF FUNCTIONAL ATROPHIC BORDER BY HELICOBACTER PYLORI INFECTION IN ATROPHIC GASTRITIS Satoru Yabe. Taiki lwahori, Taewang Nishikura, Fujio Yamagishi, Kazuyoshi Sasaki, Toshiyuki Nakajima, Kenji Fujiwara Department of Medicine 3, Saitama Medical School, Saitama JAPAN Since Helicobacter pylori (Hp) produces ammonia, an elevation of mucosul pH is expected in the Hp-infeeted stomach. If this is true, Hp infection might recess the functional atrophic border (FAB) as a results of reduction in the acid secretion area. In this study, FAB was compared between Hp-positive and negative patients using crystal violet (CV) solution as a pH indicater. (Patients and methods) Ninety-six patients were included in this study. Hp--positive group: i0 with active duodenal ulcer (DU). 11 with active gastric ulcer (GU), and 35 with atrophic gastritis (AG). Hp-.negmtive group: 31 with AG, 9 with normal subjects. Gastric juice was aspirated endoscopically for the determination of ammonia concentration. After the endoscopic atrophic border (EAB) was defined, 0.05~ CV solution was sprayed onto the gastric mucosa, and the FAB was observed. Atrophic border was divided into closed (CI,C2, C3) and open (01,02,03) types. CV solution turns green-sky bluepurplish red in the range of pll 1.0-4.0. Hp infection was determlned by both histological and serological test. (Results) (I) The ammonia concentrations (mean ± SD, gg/dl) in the gastric juice were 17.469 ± 7,768. 14,233 ± 8,032, and 12,272 ± 9,106 in DU, GU, and AG, respectively in the Hp-positive group. The concentration, 3,459 ± 1,728. was significantly lower in Hp-negative group lhan in Hp-positive group. (2) Relationship between EAB and FAB in the Hp positive and negative groups(Table).
This single centre, randomised, crossover study was conducted to investigate the effects of low doses of ranitidine on intragastric acidity. Methods: Twenty-four healthy males, mean age 23 years (range 20 to 29 years) took part in the study. Each subject took one tablet of antacid (Maalox TC TM)on the first study day and on subsequent study days received single oral doses of ranitidine 25mg, 75mg and 125mg (as the hydrochloride) and matching placebo, in accordance with a random code. The doses of study drug were administered 45 minutes after lunch and intmgastric pH was recorded for 9 hours after dosing. Each dose was separated by a two-week washout period. Results: In the first 5 hours after dosing, the decreases in integrated acidity (I-P'AUC) relative to placebo were 45.4%, 71.3% and 84.1% for ranitidine 25mg, 75rag and 125rag, respectively and -4.3% for antacid. In the 5- to 9-hour period the decreases were 20.8%, 45.0%, 65.9% and -14.2%, respectively. There was a significant linear relationship between the decrease in H + AUC and increasing ranitidine dose (p<0.0001). All doses were well tolerated.
Parameter
Post-dose Interval
Placebo
H+ AUC (mmol/L.h)
0-5 hrs 5-9 hrs
180.8 57.5
25rag
Ranitidine 75rag 125mg
92.5* 37.7*
53.5* 29.6*
24.5* 17.2" CI~3
Time pH>3 (minutes)
0-5 hrs 5-9 hrs
39.7 |13.9
58.8 120.6
1002" 148.0"*
153.5" 181.6"
* p<0.0001 and **p=0.001 for ranitidine vs placebo. Conclusion: All three maitidine doses produced significant decreases in integrated acidity compared with placebo and the decreases were dose-related. Compared with placebo, time with pH>3 was significantly greater for ranitidine 75mg and 125mg but not 25mg. No effect of antacid was observed.
PROSTAGLANDIN E2 INHIBITIS ETHANOL-INDUCED HISTAMINE RELEASE FROM ISOLATED CANINE GASTRIC MAST CELLS. K.Yakabi. A.Miyamoto and T.Nakamura. The Third Dept. of Medicine, Teikyo University School of Medicine, Chiba, Japan. Ethanol was well known to cause gastric muccsal damage in man and animal. Prostaglandin E.2(PGE2) was shown to decrease gastric mcosal damage by ethanol injection. However the mechanism by which PGE2 inhit~its ethanol-induced gastric mucosal damage was not fully understood. To elucidate the mechanism for the protection of gastric mucosa by PGE2 from ethanol-induced damage, we undertook to study the effects of ethanol and PGE2 on histamine release from isolated canine gastric mucosal mast cells. Method; Canine fundic mucosal mast ceils were dispersed and enriched to 20% by centrifugal elutriation according to SoWs method. Briefly, after blunt separation of the canine fundic mucosa from submucosa, mucosal cells were dispersed by sequential exposure ! o co/lagenase and EDTA. The mast cells were enriched by sedimentation visocity using a Beckman elutriator rotor. Mast cells were identified by toiuidine blue staining and electronmicroscopy. Finally mast cells were enriched to 20%. For study of histamine release, cells were incubated in 2ml EBSS medium with test agents(ethanoI,PGE2) for 5sin. Aliquots were centrifuged and the supematants were frozen for later histamine assay. The measurement of histamine was by radioimmunoassay using Eiken histamine assay kit. Results; Ethanol(l~20%) dose-dependently increased histamine release from isolated canie gastric mucosal mast cells. 5% ethanol caused 690% increse(P<0.O5) of histamine release and 20% ethanol caused 50times histamine relase(P<0.01) of basal group. PGE2(10"%'I0"~M) dose-dependently increased histamine release. 105M PGE2 caused 48% increase of histamine release. However, when PGE2 was added with 5% ethanol, PGE2(107~10*M)I dose-dipendently inhibited ethanot(5%)-induced histamine release. 10"TM,10~M,and 10"aM PGE2 significantly inhibited ethanol-induced histamine release by 14%(P<0.1),29%(P<0.05) and 35%(P<0.1). Conclusion;It was suggested that one of the action of PGE2 to decrease gastric mucosal damage induced by ethanol was to inhibit ethanol-induced histamine release from gastric mucosal mast cells.
DO GU AG Hp(-)
C1~3 i0 8 7 39
FAB 01~2 0 1 4 0
03 0 2 13 0
oo
o
o
o
GU AG Hp(-)
0 0 0
0 1 1
0 10 0
(Conclusion) It was suggested that Hp infection recesses functional atrophic border in AG, but not in DU and GU.
•
E X P R E S S I O N OF C Y T O K I N E m R N A IN G A S T R I C M U C O S A W I T H H E L I C O B A C T E R PYLORI INFECTION Y. Yamaoka, M. Kita, T. Kodama, N. Sawai, K. Kashima and J. Imanishi. Third Department of Internal Medicine and Department of Microbiology, Kyoto Prefectural University of Medicine, Kyoto, JAPAN Background: Helicobacterpylori(HP) infection has been considered to be a major cause of chronic gastritis and peptic ulcer disease. However, the pathogenesis of infection is still unknown. HP-associated gastritis is characterized by severe infiltration of neutrophils (PMN) and mononuclear cells (MNC) in the gastric mucosa. Cytokines have been thought to be important in accumulation and activation of these cells. Therefore, we have studied the cytokine production patterns in the mucosal biopsy specimens using the reverse transcription-PCR (RT-PCR). Methods: Total RNA was prepared from antral and corporal biopsy specimens of 32 patients, the specimens of which were analyzed for the presence of HP by culture and for the degree of gastritis by H&E stain. RT-PCR was performed using the oligonucleotideprimers specific for IL-113, IL-2, IL-3, IL-4, IL-5, IL-6, IL7, [L-8, IL-9, IL-10, TNF-ct, IFN-a, IFN-I] and IFN--I. Results: IL-6 IL-7 IL-8 IL-10 1) HP (+) gastritis Antrum 3/18 15/18 13/18 4/18 Corpus 0/17 12/17 10/17 1/17 2) HP (-) gastritis Antrum 0/8 3/8 3/8 0/8 Corpus 0/9 1/9 1/9 0/9 3) HP (-) normal Antrum 0/6 2/6 0/6 0/6 Corpus 0/6 1/6 0/6 0/6 Antrum: l)vs3) IL-7 (p =0.038), IL-8 (p =0.003) l)vs2) IL-7 (p ---0.032) Corpus: l)vs3)IL-7(p=0.035),IL-8(p=0.017) l)vs2)IL-7(p=0.006) IL-8 (p =0.024) Although IL-I~ and IFN-v mRNA were detected in the majority of specimens, IL-2, IL-3, IL-4, IL-5, IL-9, TNF-ct and IFN-I~ mRNA were not detected at all. IL-6 mRNA was detected in 0nly 3 specimens from HP(+) gastritis with severe infiltration of PMN and MNC. IL-10 mRNA was also detected in only the specimens from HP(+) gastritis with severe infiltration of PMN. The expressions of IL-7 and IL-8 mRNA were significantly higher in HP(+) gastritis than in HP(-) normal controls. There was a significant correlation between the IL-8 mRNA expression and the severity of gastritis both in the antrum and COrpUS. On the other hand, there was a significant correlation between the IL-7 mRNA expression and the severity of gastritis only in the antrum. Conclusion: Many kinds of cytokine mRNA can be detected in the mucosal biopsy specimens using the RT-PCR. In addition, some cytokines, especially IL-7 and IL-8, play important roles in HPassociated gastritis.
Esophageal, Gastric, and Duodenal Disorders A261
• DECREASE OF INTRAGASTRIC ACIDITY FOLLOWING LOW DOSES OF RANITIDINE JW Wyeth 1, RE_ Pounder 1, CCL Snell2, IRoyal Free Hospital School of Medicine, London, ~;-----~GlaxoResearch & Development Ltd, Grcenford, UK
REGRESSION OF FUNCTIONAL ATROPHIC BORDER BY HELICOBACTER PYLORI INFECTION IN ATROPHIC GASTRITIS Satoru Yabe. Taiki lwahori, Taewang Nishikura, Fujio Yamagishi, Kazuyoshi Sasaki, Toshiyuki Nakajima, Kenji Fujiwara Department of Medicine 3, Saitama Medical School, Saitama JAPAN Since Helicobacter pylori (Hp) produces ammonia, an elevation of mucosul pH is expected in the Hp-infeeted stomach. If this is true, Hp infection might recess the functional atrophic border (FAB) as a results of reduction in the acid secretion area. In this study, FAB was compared between Hp-positive and negative patients using crystal violet (CV) solution as a pH indicater. (Patients and methods) Ninety-six patients were included in this study. Hp--positive group: i0 with active duodenal ulcer (DU). 11 with active gastric ulcer (GU), and 35 with atrophic gastritis (AG). Hp-.negmtive group: 31 with AG, 9 with normal subjects. Gastric juice was aspirated endoscopically for the determination of ammonia concentration. After the endoscopic atrophic border (EAB) was defined, 0.05~ CV solution was sprayed onto the gastric mucosa, and the FAB was observed. Atrophic border was divided into closed (CI,C2, C3) and open (01,02,03) types. CV solution turns green-sky bluepurplish red in the range of pll 1.0-4.0. Hp infection was determlned by both histological and serological test. (Results) (I) The ammonia concentrations (mean ± SD, gg/dl) in the gastric juice were 17.469 ± 7,768. 14,233 ± 8,032, and 12,272 ± 9,106 in DU, GU, and AG, respectively in the Hp-positive group. The concentration, 3,459 ± 1,728. was significantly lower in Hp-negative group lhan in Hp-positive group. (2) Relationship between EAB and FAB in the Hp positive and negative groups(Table).
This single centre, randomised, crossover study was conducted to investigate the effects of low doses of ranitidine on intragastric acidity. Methods: Twenty-four healthy males, mean age 23 years (range 20 to 29 years) took part in the study. Each subject took one tablet of antacid (Maalox TC TM)on the first study day and on subsequent study days received single oral doses of ranitidine 25mg, 75mg and 125mg (as the hydrochloride) and matching placebo, in accordance with a random code. The doses of study drug were administered 45 minutes after lunch and intmgastric pH was recorded for 9 hours after dosing. Each dose was separated by a two-week washout period. Results: In the first 5 hours after dosing, the decreases in integrated acidity (I-P'AUC) relative to placebo were 45.4%, 71.3% and 84.1% for ranitidine 25mg, 75rag and 125rag, respectively and -4.3% for antacid. In the 5- to 9-hour period the decreases were 20.8%, 45.0%, 65.9% and -14.2%, respectively. There was a significant linear relationship between the decrease in H + AUC and increasing ranitidine dose (p<0.0001). All doses were well tolerated.
Parameter
Post-dose Interval
Placebo
H+ AUC (mmol/L.h)
0-5 hrs 5-9 hrs
180.8 57.5
25rag
Ranitidine 75rag 125mg
92.5* 37.7*
53.5* 29.6*
24.5* 17.2" CI~3
Time pH>3 (minutes)
0-5 hrs 5-9 hrs
39.7 |13.9
58.8 120.6
1002" 148.0"*
153.5" 181.6"
* p<0.0001 and **p=0.001 for ranitidine vs placebo. Conclusion: All three maitidine doses produced significant decreases in integrated acidity compared with placebo and the decreases were dose-related. Compared with placebo, time with pH>3 was significantly greater for ranitidine 75mg and 125mg but not 25mg. No effect of antacid was observed.
PROSTAGLANDIN E2 INHIBITIS ETHANOL-INDUCED HISTAMINE RELEASE FROM ISOLATED CANINE GASTRIC MAST CELLS. K.Yakabi. A.Miyamoto and T.Nakamura. The Third Dept. of Medicine, Teikyo University School of Medicine, Chiba, Japan. Ethanol was well known to cause gastric muccsal damage in man and animal. Prostaglandin E.2(PGE2) was shown to decrease gastric mcosal damage by ethanol injection. However the mechanism by which PGE2 inhit~its ethanol-induced gastric mucosal damage was not fully understood. To elucidate the mechanism for the protection of gastric mucosa by PGE2 from ethanol-induced damage, we undertook to study the effects of ethanol and PGE2 on histamine release from isolated canine gastric mucosal mast cells. Method; Canine fundic mucosal mast ceils were dispersed and enriched to 20% by centrifugal elutriation according to SoWs method. Briefly, after blunt separation of the canine fundic mucosa from submucosa, mucosal cells were dispersed by sequential exposure ! o co/lagenase and EDTA. The mast cells were enriched by sedimentation visocity using a Beckman elutriator rotor. Mast cells were identified by toiuidine blue staining and electronmicroscopy. Finally mast cells were enriched to 20%. For study of histamine release, cells were incubated in 2ml EBSS medium with test agents(ethanoI,PGE2) for 5sin. Aliquots were centrifuged and the supematants were frozen for later histamine assay. The measurement of histamine was by radioimmunoassay using Eiken histamine assay kit. Results; Ethanol(l~20%) dose-dependently increased histamine release from isolated canie gastric mucosal mast cells. 5% ethanol caused 690% increse(P<0.O5) of histamine release and 20% ethanol caused 50times histamine relase(P<0.01) of basal group. PGE2(10"%'I0"~M) dose-dependently increased histamine release. 105M PGE2 caused 48% increase of histamine release. However, when PGE2 was added with 5% ethanol, PGE2(107~10*M)I dose-dipendently inhibited ethanot(5%)-induced histamine release. 10"TM,10~M,and 10"aM PGE2 significantly inhibited ethanol-induced histamine release by 14%(P<0.1),29%(P<0.05) and 35%(P<0.1). Conclusion;It was suggested that one of the action of PGE2 to decrease gastric mucosal damage induced by ethanol was to inhibit ethanol-induced histamine release from gastric mucosal mast cells.
DO GU AG Hp(-)
C1~3 i0 8 7 39
FAB 01~2 0 1 4 0
03 0 2 13 0
oo
o
o
o
GU AG Hp(-)
0 0 0
0 1 1
0 10 0
(Conclusion) It was suggested that Hp infection recesses functional atrophic border in AG, but not in DU and GU.
•
E X P R E S S I O N OF C Y T O K I N E m R N A IN G A S T R I C M U C O S A W I T H H E L I C O B A C T E R PYLORI INFECTION Y. Yamaoka, M. Kita, T. Kodama, N. Sawai, K. Kashima and J. Imanishi. Third Department of Internal Medicine and Department of Microbiology, Kyoto Prefectural University of Medicine, Kyoto, JAPAN Background: Helicobacterpylori(HP) infection has been considered to be a major cause of chronic gastritis and peptic ulcer disease. However, the pathogenesis of infection is still unknown. HP-associated gastritis is characterized by severe infiltration of neutrophils (PMN) and mononuclear cells (MNC) in the gastric mucosa. Cytokines have been thought to be important in accumulation and activation of these cells. Therefore, we have studied the cytokine production patterns in the mucosal biopsy specimens using the reverse transcription-PCR (RT-PCR). Methods: Total RNA was prepared from antral and corporal biopsy specimens of 32 patients, the specimens of which were analyzed for the presence of HP by culture and for the degree of gastritis by H&E stain. RT-PCR was performed using the oligonucleotideprimers specific for IL-113, IL-2, IL-3, IL-4, IL-5, IL-6, IL7, [L-8, IL-9, IL-10, TNF-ct, IFN-a, IFN-I] and IFN--I. Results: IL-6 IL-7 IL-8 IL-10 1) HP (+) gastritis Antrum 3/18 15/18 13/18 4/18 Corpus 0/17 12/17 10/17 1/17 2) HP (-) gastritis Antrum 0/8 3/8 3/8 0/8 Corpus 0/9 1/9 1/9 0/9 3) HP (-) normal Antrum 0/6 2/6 0/6 0/6 Corpus 0/6 1/6 0/6 0/6 Antrum: l)vs3) IL-7 (p =0.038), IL-8 (p =0.003) l)vs2) IL-7 (p ---0.032) Corpus: l)vs3)IL-7(p=0.035),IL-8(p=0.017) l)vs2)IL-7(p=0.006) IL-8 (p =0.024) Although IL-I~ and IFN-v mRNA were detected in the majority of specimens, IL-2, IL-3, IL-4, IL-5, IL-9, TNF-ct and IFN-I~ mRNA were not detected at all. IL-6 mRNA was detected in 0nly 3 specimens from HP(+) gastritis with severe infiltration of PMN and MNC. IL-10 mRNA was also detected in only the specimens from HP(+) gastritis with severe infiltration of PMN. The expressions of IL-7 and IL-8 mRNA were significantly higher in HP(+) gastritis than in HP(-) normal controls. There was a significant correlation between the IL-8 mRNA expression and the severity of gastritis both in the antrum and COrpUS. On the other hand, there was a significant correlation between the IL-7 mRNA expression and the severity of gastritis only in the antrum. Conclusion: Many kinds of cytokine mRNA can be detected in the mucosal biopsy specimens using the RT-PCR. In addition, some cytokines, especially IL-7 and IL-8, play important roles in HPassociated gastritis.