The effect of compound 48/80 and of electrical field stimulation on mast cells in the isolated mouse stomach

It has been shown that compound 48/80 evokes acid secretion in the isolated mouse stomach by releasing histamine from mast cells. The aim of the present work was to study the distribution and type of mast cells in the isolated mouse stomach and to examine the effect of compound 48/80 and of electrical field stimulation (EFS) on these cells.Histological examination of the stomachs showed only the presence of connective tissue mast cells (CTMC) in all parts of the stomach except in the glandular mucosa where mucosal mast cells (MMC) predominated (MMC–71%, CTMC–29%). Compound 48/80 and EFS did not affect MMC, whereas CTMC showed marked degranulation in all parts of the stomach including the glandular mucosa. It can be concluded that CTMC, present in the gastric mucosa and being sensitive also to EFS, may be involved in the regulation of gastric secretion in the mouse.     

Development of different mast cell types in the opossum<Emphasis Type="Italic"> Didelphis albiventris</Emphasis>

Previous studies have disclosed three types of mast cell in opossums: connective tissue (CTMC), mucosal (MMC), and lymphatic sinus (LSMC). In contrast to most opossum lymph nodes, the mesenteric lymph node is virtually devoid of LSMC, displaying medullary cord CTMC. The present study aimed to describe the development of these mast cell populations. Toluidine blue staining and a histochemical method for demonstrating heparin allowed the identification of immature and mature mast cells. Immature CTMC devoid of detectable heparin were rare until postnatal day 10. Mature CTMC filled with heparin-containing granules became numerous by day 30 to day 40. In the ileum, despite the presence of mature CTMC in the submucosa and mucosa (villus base), immature mast cells first appeared in the villus core by day 65 and adult features were apparent by day 100. In LSMC-containing lymph nodes, immature mast cells were found in lymphatic sinuses by day 10. Clear signs of LSMC differentiation were observed from day 20. Compared with the 10-day value, the mean diameter of cytoplasmic granules at day 40 had doubled and that at day 110 had tripled. In the mesenteric lymph nodes, immature mast cells differentiated into lymphatic sinus CTMC-like cells. After day 80, most of them were located in medullary cords. Weaning and complete maturation of mucosa preceded the differentiation of MMC. In lymph nodes, LSMC differentiation occurred in parallel with the development of the medullary region and deep cortex units.     

Development of different mast cell types in the opossum Didelphis albiventris

Previous studies have disclosed three types of mast cell in opossums: connective tissue (CTMC), mucosal (MMC), and lymphatic sinus (LSMC). In contrast to most opossum lymph nodes, the mesenteric lymph node is virtually devoid of LSMC, displaying medullary cord CTMC. The present study aimed to describe the development of these mast cell populations. Toluidine blue staining and a histochemical method for demonstrating heparin allowed the identification of immature and mature mast cells. Immature CTMC devoid of detectable heparin were rare until postnatal day 10. Mature CTMC filled with heparin-containing granules became numerous by day 30 to day 40. In the ileum, despite the presence of mature CTMC in the submucosa and mucosa (villus base), immature mast cells first appeared in the villus core by day 65 and adult features were apparent by day 100. In LSMC-containing lymph nodes, immature mast cells were found in lymphatic sinuses by day 10. Clear signs of LSMC differentiation were observed from day 20. Compared with the 10-day value, the mean diameter of cytoplasmic granules at day 40 had doubled and that at day 110 had tripled. In the mesenteric lymph nodes, immature mast cells differentiated into lymphatic sinus CTMC-like cells. After day 80, most of them were located in medullary cords. Weaning and complete maturation of mucosa preceded the differentiation of MMC. In lymph nodes, LSMC differentiation occurred in parallel with the development of the medullary region and deep cortex units.     

人胃粘膜肥大细胞的分布与超微结构

光镜观察了２５例人胃底部组织甲苯胺蓝染色切片的粘膜肥大细胞的分布。电镜观察了５例人胃底活检材料内粘膜肥大细胞的超微结构。     

Gut mucosal mast cells in Nippostrongylus-primed rats are the major source of secreted rat mast cell protease II following systemic anaphylaxis

The distribution of the predominant chymotrypsin-like enzyme of mucosal mast cells (rat mast cell protease II: RMCP II) was examined in naive and Nippostrongylus-primed rats both before and after the induction of systemic anaphylaxis. Anaphylactic secretion of RMCP II following i.v. challenge of primed rats with worm antigen was accompanied by significant depletion of this enzyme from the jejunal and gastric mucosae; the concentrations were not altered in the ileum and colon. Despite significant increases in the levels of RMCP II in lung and mesenteric lymph node following infection with N. brasiliensis there was no anaphylactic depletion of this enzyme from these sites. No RMCP II was detected in liver, spleen, kidney or bone marrow either before or after systemic anaphylaxis. Mucosal mast cells were depleted from the jejunal, gastric and colonic mucosae following antigen challenge of primed rats. These data provide further evidence that gastrointestinal mucosal mast cells are the major source of secreted RMCP II following systemic anaphylaxis in the rat.     

Effect of Amylin on Mast Cell Secretion as a Possible Mechanism Increasing Gastric Mucosa Resistance

Water-immersion restraint stress increased secretory activity of mast cells and led to the formation of erosive lesions in the gastric mucosa. Intraperitoneal administration of amylin in a dose of 0.5 g/kg 1 h before stress suppressed degranulation of mast cells and decreased the severity of gastric mucosa damages. In in vitro experiments amylin abolished the activating effects of acetylcholine and bradykinin on mast cell degranulation. Amylin-induced stabilization of activated mast cells probably underlies its protective effects during ulceration.     

Effects of a mast cell stabiliser (FPL 52694) on human gastric secretion

The effects of a new monochrome mast cell stabilizer (FPL 52694) on gastric secretion have been studied in healthy volunteers. When administered orally for 3 days, FPL 52694 consistently reduced nocturnal and pentagastrin-stimulated secretion of acid by about 50%. Mealstimulated gastric secretion was not affected by the drug. Inhibition of the gastric secretory response to pentagastrin was not significant when single doses of the drug were administered either into the stomach or the duodenum, but administration of multiple doses resulted in significant inhibition. The drug does not inhibit gastric secretion by injuring the gastric mucosa, since the gastric mucosal barrier to back diffusion of hydrogen ions is not affected. We conclude that this mast cell stabilizer provides an interesting tool for studying aspects of the physiological control of gastric secretion and may have a therapeutic role in peptic ulceration.     

Inhibition of Vagally Stimulated Gastric Secretion after Mucosal Mast Cell Degranulation

Total vagotomy was carried out on rats through laparotomy. and the peripheral end of the vagus was electrically stimulated for one hour after closing the ventricle with a Shay ligature. after the stimulation, the gastric fluid was collected, its volume, pH and concentration of free HCI were measured. Free HCl was secreted significantly less in the ventricles of rats whose mucosal mast cells had been degranulated by dexamethasone before the vagus stimulation, than in the rats not treated with dexamethasone. The importance of the synthesis and accumulation of histamine in the mast cells of the gnstrir mucosa as a peripheral link in the stimulatory chain of gastric secretion is discussed.     

Stoffwechsel und physiologische Funktion von Histamin im Magen

Zusammenfassung 1. Histamin kommt in der Magenwand aller untersuchten Arten in hoher Konzentration vor, in der Schleimhaut vor allem in oder nahe bei Belegzellen, mucoiden Drüsenzellen, Mastzellen und enterochromaffinen Zellen.2. Histamin wird in der Magenschleimhaut aller untersuchten Arten in ausreichendem Maße vor allem durch die spezifische Histidindecarboxylase gebildet, die sich in erster Linie in oder in der Nähe von den Belegzellen und mucoiden Drüsenzellen befindet. Die unspezifische HDC kommt in der Mucosa vor allem in den enterochromaffinen Zellen vor.3. Die Histamininaktivierung im Magengewebe erfolgt bei Mensch, Katze, Kaninchen, Meerschweinchen und Maus überwiegend durch Histaminmethyltransferase, bei der Ratte durch Diaminoxydase.4. Histamin stimuliert die Magensaftsekretion bei allen untersuchten Arten. Zwischen Histamingehalt der Schleimhaut, Histaminbildung und Empfindlichkeit der Belegzellen gegenüber Histamin besteht eine Korrelation.5. Histamin wird durch eine große Zahl von Substanzen aus Mastzell- und Nichtmastzellspeichern freigesetzt. Beweisgründe werden erbracht, daß Gastrin und cholinerge Mechanismen die Magensaftsekretion durch Freisetzung von Histamin stimulieren. Histamin ist danach der finale Chemostimulator.6. Durch Verdauungsvorgänge, Gastrin und Anlegen eines portocavalen shunts kommt es zu vermehrter Histaminbildung durch Induktion der spez. HDC, ebenso durch Glucocorticoide, Thyroxin und entzündungshemmende Substanzen wie Indomethazin, Salicylate, Phenylbutazon, Fluphenaminsäure. Auch Streß, Kältereiz und Serotonin haben diese Wirkung.7. Die Wirkung der Antihistaminica auf die Magensaftsekretion ist noch nicht befriedigend geklärt. Die Hemmung der Magensaftsekretion bei der Ratte durch das Antihistaminicum Phloxin ist sicher spezifisch.8. Bei der Ratte ist die physiologische Wirkung von Histamin als finalem Chemostimulator der Magensekretion erwiesen, bei allen anderen Species wahrscheinlich.

---

Summary 1. The stomach of men and all of the animals, which have been investigated, contains histamine in a high concentration. In the mucosa histamine is found especially in or near the parietal cells (fundus and corpus), by the mucoid cells (cardia and pyloric glands, mucous neck cells), but also in the mast cells and enterochromaffine cells. In the submucosa and muscularis propria histamine could be demonstrated in mast cells, muscle fibers of the smooth muscle and arterioles by autoradiography and fluorescence microscopy. Further its localisation is suggested in the wall of capillaries and in nerves. So a great number of cellular pools for histamine in the gastric wall must be postulated. It is possible that histamine changes its position between these pools.2. In the stomach of men and all of the animals which have been investigated, histamine is formed in a sufficient amount by the specific histidine decarboxylase. These results have been obtained by isotopic and nonisotopic assays. The highest activity of the specific HDC mostly is demonstrated in the fundus, the lowest in the pylorus. The enzyme is found in or near by parietal cells and mucoid cells, its localisation in mast cells and endothel cells is suggested. The activity of the unspecific HDC is fairly the same in all regions of gastric mucosa. The enzyme is found in the enterochromaffine cells, but is suggested to be in mast cells and nerves, too.3. In man, cat, rabbit, guinea pig and mice histamine in the gastric tissue is inactivated especially by histamine methyltransferase, by diamine oxidase especially in the rat. The formation of histamine nucleotide, acetylation and transamination may be important too.4. Histamine stimulates the gastric secretion in men and all of the animals which have been investigated, with exception of lower amphibians and elasmobranch fishes which possibly have got a dose of histamine, which has not been sufficient for stimulation. Histamine in every way of application stimulates the gastric secretion, but applicated orally, high amounts (50–100 mg) are needed. A correlation exists between the histamine content, the formation of histamine and the sensitivity of the parietal cells with respect to histamine stimulation. The gastric juice after histamine stimulation shows a high acidity and a great volumen of juice, but the pepsin and mucin content is low in many species.5. From mast cells compound 48/80, d-tubocurarine and antihistaminics liberate histamine, from nonmast cell storages, especially in the gastric mucosa, gastrin and parasympathicomimetic mechanisms stimulate gastric secretion by liberation of histamine. Histamine is the final chemostimulator. The importance of liberation and formation of histamine in striatic muscles during chronically elevated muscle tonus for development of high basis secretion and duodenal ulcer is discussed.6. During digestion, by gastrin injection and portocaval shunt histamine formation is elevated by induction of the specific HDC. Hormonal influences like injection of glucocorticoids, thyroxine, and antiinflammatory drugs, for instance indomethazine, salicylic acid, phenylbutazone and fluphenamic acid stimulate histamine formation in the same way, also stress by restraint and cold or serotonin. Glucocorticoids inhibit the histamine formation in all organs and tissues except the stomach.7. The investigation of the action of antihistaminics on gastric secretion today has not still obtained satisfactoring results. Parenterally applicated the antihistaminics have been found to be ineffective in all species except the pigeon. By local or i.a. application immediately before the stomach they inhibit the gastric secretion. In the dog, parenterally applicated they are effective, if tween 20 simultaneously is injected. The question of specific or unspecific action of antihistaminics (by mucosal injury or by cholinolytic action) appears to be not completely cleared, especially because in salivary glands of dogs it has been shown that antihistaminics injected immediately before the gland inhibit the secretion of saliva in a specific way. Phloxine inhibits the gastric secretion in the rat specifically.8. In the rat complete evidence is presentated, that histamine is the physiological final chemostimulator of the gastric secretion. In men and all of the other investigated animals evidence is strongly supported, but not completed, that histamine acts on the gastric tissue in the same way. The pecularities of the rat with respect to the metabolism of histamine and gastric secretion should not be exaggerated.

---

---

Unserem verehrten Lehrer, Herrn Prof. Dr. Dr.E. Werle, zum 65. Geburtstag in Dankbarkeit gewidmet.     

抑郁模型大鼠胃窦部变态反应及超微结构改变

目的：观察抑郁模型大鼠胃窦部变态反应及超微结构改变。方法:将慢性、不可预见性、非同质性应激原作为抑郁模型,通过免疫荧光组织化学及超微结构形态学观察抑郁模型组(n=10)和正常对照组 (n=10)大鼠胃窦部的免疫变态反应及形态学的变化。结果:抑郁模型组大鼠胃窦黏膜组织肥大细胞胰蛋白酶1（MCP-1）平均免疫荧光强度值(37.4±7.7), 显著高于对照组（24.8±5.6）,P＜0.05。抑郁模型大鼠胃窦黏膜组织肥大细胞激活、增殖及颗粒增生。超微电镜观察发现,抑郁模型组大鼠胃窦黏膜组织肥大细胞表现为：肥大细胞向血管、神经和单核细胞周围渗透,肥大细胞膜折皱,细胞内含有非同质的、非均匀、低电子密度颗粒,部分颗粒到达膜表面、成为丝状物、空颗粒及脂质体,细胞核呈梭形;肥大细胞周围白细胞、单核细胞、巨噬细胞渗出;肥大细胞激活脱颗粒,脱落的颗粒趋化到巨噬细胞表面;白细胞颗粒与肥大细胞膜黏附形成桥接;巨噬细胞吞噬肥大细胞激活脱落的颗粒。结论:抑郁模型大鼠胃窦部发生免疫变态反应,肥大细胞对微环境的变化一方面表现肥大细胞细胞数显著增加,另一方面表现为其表型的变化。     

Effect of ketotifen and oxatomide on histamine secretion from mast cells

The anti-histaminic drugs ketotifen and oxatomide have a dual effect on rat peritoneal mast cells. At high concentrations they induce histamine release, whereas at low concentrations they inhibit secretion evoked by IgE-directed ligands. The latter effect is observed in the presence and absence of exogenous calcium. The significance of this result for the general mode of action of anti-anaphylactic drugs is discussed. Neither compound liberates histamine from isolated mesenteric cells from the rat or guinea pig, further emphasizing the functional heterogeneity of mast cells from different sources.     

Mast cells from human gastric mucosa: A comparative study with lung and colonic mast cells

Mast cells isolated from human gastric mucosa released histamine on challenge with IgE-directed ligands and calcium ionophores but were esentially unresponsive to a variety of non-immunological stimuli. Moreover, immunologically induced histamine secretion from these cells was inhibited by a number of anti-allergic agents including anti-asthmatic chromones, -adrenoceptor agonists and phosphodiesterase inhibitors. In total, these data indicate that mast cells from the human gastric mucosa are in many respects functionally similar to their lung and colonic counterparts.     

Protection of the Gastric Mucosa against the Lesions Caused by Reserpine through Degranulaton of Mucosal Mast Cells

The ulcerating effects of reserpinisation in the gastric mucosa of rats were studied by special observation of the function of the gastric mucosal mast cells and the changes in mitotic activity of the epithelial cells. The significance of degranulation of the mucosal mast cells for the ulcerating effect of reserpine was also studied after the pretreatment of rats with dexamethasone. Immediately after the dexamethasone treatment there was a pronounced decrease in mitotic count of the gastric mucosal epithelium which 2—3 days later when the mast cells of the gastric mocusa were almost completely degranulated was followed by a 2—3 fold increase in mitotic activity as compared with controls. 10 and 20 days later the mitotic count in the gastric mucosal epithelium decreased below the controls values, the mucosal mast cells being re-granulated over the control values. The changes in mitotic count in the epithelium of duodenum. ileum and colon under the same conditions were scanty. The degranulation of the mast cells was markedly slighter in the intestinal than in the gastric mucosa. In the reserpinized rats the number of mitoses in the gastric epithelium decreased simultaneously with the degranulation of the mast cells. The degranulation of the mast cells in the duodenal mucosa was slighter and no changes in mitotic activity occurred in the duodenal epithelium. Lesions appeared during reserpinisation in the ventricular mucosa. Reserpinisation after the dexamethasone treatment did not cause ulcer formation in the gastric mucosa, nor did it in this phase cause inhibition in the mitotic activity, typical of which was an overshooting 2—3 days after dexamethasone treatment. The mechanism of gastric mucosal ulceration is discussed.     

Histamine release from mast cells of various species induced by histamine releasing factor from human lymphocytes

The aim of our work was to investigate the effect of histamine releasing factor (HRF), producedin vitro by lymphocytes obtained from atopic and non-atopic asthmatics, on mast cells of various species (mouse—peritoneal mast cells, hamster and rat—peritoneal and pleural mast cells, guinea-pig — mesenteric and pulmonary mast cells). We found that human HRF is able to release histamine from the examined mast cell populations in a dose-dependent fashion. Mast cells from various species differed in their susceptibility to the action of HRF; rat pleural and guinea-pig mesenteric and pulmonary mast cells were the most susceptible, while mouse and hamster peritoneal mast cells —the least susceptible. The presence of 50% D₂O in the medium significantly increased HRF-induced histamine release from rat mast cells, while the addition of phosphatidylserine did not change it. HRF-induced histamine release from guinea-pig mesenteric mast cells was not related to sensitization of these cells.We also compared histamine release from guinea-pig pulmonary and mesenteric mast cells induced by human HRF producedin vitro by lymphocytes obtained from atopic and non-atopic asthmatics. We have found that supernatant from atopic asthmatics lymphocyte cultures released significantly more histamine than supernatant from non-atopic asthmatics lymphocyte cultures.Our studies give evidence that human HRF acts across the species barrier and induces histamine release from mast cells of various species. The mechanism of HRF action on mast cells seems to be different from that of allergen.This work was supported by Polish academy of Sciences (Grant CPBP 06.01).     

Stimulation of afferent nerve endings by intragastric capsaicin protects against ethanol-induced damage of gastric mucosa

Ablation of capsaicin-sensitive afferent neurons enhances experimentally induced ulceration in the rat gastric mucosa, which suggests that these neurons are involved in gastric mucosal protection. To provide direct evidence for such a function it was investigated whether stimulation of afferent nerve endings by the intragastric administration of capsaicin could counteract the ulcerogenic effect of 25% ethanol. Capsaicin (3.2-640 microM), administered together with ethanol, inhibited the development of haemorrhagic lesions in a concentration-dependent fashion but did not alter the ethanol-induced fall in the gastric potential difference. This suggests that capsaicin does not prevent ethanol from damaging gastric epithelial cells but can counteract the vascular lesions caused by ethanol. The anti-lesion effect of intragastric capsaicin was absent in adult rats which had been treated with a high dose of systemic capsaicin as neonates in order to achieve a permanent degeneration of unmyelinated afferent neurons. It would appear, therefore, that intragastric capsaicin reduces lesion formation by an action on afferent neurons. The protective effect of intragastric capsaicin was not altered following acute subdiaphragmatic vagotomy, acute removal of the coeliac-superior mesenteric ganglion complex, acute bilateral ligation of the adrenal glands, or pretreatment of the rats with atropine or guanethidine. These findings indicate that stimulation of afferent neurons by intragastric capsaicin affords protection of the rat gastric mucosa against ethanol-induced damage. As the autonomic nervous system is not involved gastroprotection appears to represent a local effector function of sensory nerve endings in the stomach.     

Mast cells and eosinophils as regulators of functional homeostasis in the gastric mucosa

The numerous mast cells of the interstitial tissue of the gastric mucosa are the sites of histamine synthesis and storage. If this biogenic amine is liberated from these cells it transmits stimulation of gastric acid secretion to the epithelial cells. However, the effect of this released histamine is limited by the eosinophilic leukocytes, which find their way to the origin of histamine and decompose it. The functional level of these two kinds of cells is controlled by the neuroendocrine system.     

Isolation and some properties of mast cells from the mesentery of the rat and guinea pig

A number of enzymes were screened for their ability to dissociate mesenteric tissues from the rat and guinea pig into their component cells. The bacterial enzyme collagenase was found to be the most satisfactory agent and a procedure based on the use of this protease was developed. The resulting suspensions contained 1–2% free mast cells and exhibited a low (ca. 5%) spontaneous release of histamine. The tissue cells contained less histamine than rat peritoneal mast cells and the guinea pig cells were smaller in size. Cells obtained from actively sensitized animals responded to antigenic challenge more strongly than the chopped tissue indicating that they were functionally intact. Rat mesenteric cells could be passively sensitized with homologous reaginic antibody and also responded to anti-rat IgE. The immunologically induced releases from rat mesenteric and peritoneal cells showed differing sensitivities to potentiation by phosphatidyl serine but the responses were directly comparable in the absence of this effect. Rat mesenteric cells also responded, but less effectively than the peritoneal cells, to the ionophore A23187, concanavalin A, ATP and basic secretagogues. They were, however, essentially refractory to the action of dextran. In contrast, guinea pig mast cells responded strongly only to the ionophore and weakly or not at all to the other agents. These results indicate marked inter-and intra-species differences in the reactivity of mast cells and suggest that rat peritoneal cells should not be used as the sole model for studying histamine secretion.     

Effect of TRPV1 on Activity of Isoforms of Constitutive Nitric Oxide Synthase during Regulation of Bicarbonate Secretion in the Stomach

Bulletin of Experimental Biology and Medicine - Application of mild irritants (1 M NaCl; pH 2.0) on the gastric mucosa potentiates the protective secretion of bicarbonates by epithelial cells. This...     

Histamine secretion from mast cells treated with chlortetracycline (aureomycin): a novel calcium ionophore

The novel calcium ionophore chlortetracycline (CTC) induced histamine secretion (≤90%) from isolated rat peritoneal mast cells in a pH and dose-dependent fashion. The process was dependent on exogenous calcium ions and was inhibited by extremes of temperature and metabolic blockers. The release was rapid, being essentially complete within 1 min, but the half-life of the process varied inversely with the concentration of the ionophore. In contrast to receptor-mediated ligands, but in keeping with other ionophores, the activated state induced by CTC did not decay with time. The secretion was effectively inhibited, according to the concentration of the ionophore, by disodium cromoglycate and other anti-allergic or cyclic AMP-active drugs. These results confirm our previous contention that these agents do not act on receptor-mediated calcium-channels. CTC induced a significant (≤50%) release of histamine from enzymically dispersed rat mesenteric mast cells but was essentially inactive against isolated mast cells from the mesentery or lung of the guinea-pig. These results extend our former observations on the functional heterogeneity of mast cells and show that, in common with other secretagogues, ionophores may exhibit selectivity in their mode of action.