Histamine storage and formation in canine gastric mucosa--effect of pentagastrin stimulation

Histamine storage and formation in the dog gastric mucosa were studied during basal conditions and after pentagastrin stimulation. Histamine formation (histidine decarboxylase activity), histamine content as well as the density of mast cells of the oxyntic gland mucosa were evenly distributed. Histamine content of the mucosa was significantly correlated to the density of mucosal mast cells. In the basal secretory state, histamine formation and histamine content of the oxyntic gland mucosa were of the same magnitude as in the antral mucosa. Pentagastrin stimulation induced a small but significant decrease in histamine content of the oxyntic gland mucosa and a subsequent acceleration in the rate of amine formation. Neither histamine content nor histidine decarboxylase activity of the antral mucosa was affected by pentagastrin infusion.     

新生小鼠胃内组胺免疫反应细胞的形态及分布

目的：观察出生后小鼠胃内组胺免疫反应细胞的个体发生、分布、形态及数量变化。方法：免疫组织化学技术。结果：小鼠出生后第5天,胃体部粘膜上皮中出现组胺阳性细胞,此后随着胃体部粘膜的发育,组胺阳性细胞数量明显增多,密集分布于胃体部粘膜下1/3处的上皮内。胃粘膜下层中也可见少量组胺弱阳性细胞。上皮内的组胺阳性细胞多为闭合型,胞体较小,常聚集、环抱壁细胞。结论：小鼠胃体部粘膜中组胺阳性细胞出现的时间较G细胞、D细胞、EC细胞晚,随着小鼠的生长发育,其数量呈显著性增加。位于胃粘膜下1/3处上二皮内的组胺阳性细胞可能为肠嗜铬样细胞（ECL细胞）,ECL细胞释放的组胺,有可能通过旁分泌的方式作用于壁细胞。     

Histamine metabolism of the guinea-pig gastric mucosa.

1. The mobilization of gastric mucosal histamine as reflected by changes in formation and content has been studied in guinea-pigs on feeding and after injections of pentagastrin or 2-deoxy-D-glucose. 2. Re-feeding fasting guinea-pigs as well as injections of pentagastrin or 2-deoxy-D-glucose raised the rate of mucosal histamine formation; pentagastrin induced a fourfold rise. 3. Some properties of the enzyme catalysing the formation of histamine were examined. The results indicated that this enzyme is histidine decarboxylase, L-histidine carboxy-lyase, E.C. 4.1.1. 22. 4. The enzyme imidazole-N-methyl transferase, E.C. 2.1.1.8, which carries out methylation of the imidazole ring to yield 1-methyl-4-(beta-aminoethyl)imidazole (methylhistamine), was found in high amounts in the mucosa. The enzyme did not change upon stimulation of the mucosa. 5. The metabolism of histamine within the gastric mucosa is discussed in relationship to a suggested role of the amine in exciting acid secretion.     

Expression of non‐mast cell histidine decarboxylase in tumor‐associated microvessels in human esophageal squamous cell carcinomas

Histamine is produced by mast cells and many other types of cells. The role of histamine released from mast cells in promoting tumor angiogenesis has been intensively studied; however, the role of non‐mast cell histamine in regulating tumor angiogenesis has been largely ignored. In this study, tissue specimen sections from 43 patients with esophageal squamous cell carcinoma (ESCC) and normal esophageal biopsies from 17 heath individuals obtained from a high incidence area of north China were used to assess changes in microvessel density (MVD) and non‐mast cell L‐histidine decarboxylase (HDC) (the only rate‐limiting enzyme that catalyzes the formation of histamine from L‐histidine) expression in the tumor microenvironment by immunohistochemistry (IHC). In addition, the cellular characterization of non‐mast cell HDC‐positive cells in microvessels was examined by double IHC combined with HDC/CD34 and HDC/PCNA antibodies. These IHC analyses revealed a significantly increased HDC‐positive MVD in ESCC as compared with normal controls, which accounted for ～61% of CD34‐labeled general MVD in ESCC. Furthermore, IHC in serial sections and double IHC showed that most of these HDC‐positive cells were CD34‐positive endothelial cells in microvessels with an increased proliferative capacity. Thus, our results suggest that non‐mast cell histamine expressed in endothelial cells of microvessels could be an additional cellular source and might play a role in regulating angiogenesis in ESCC.     

Changes in protein kinase C activity during histamine release from activated rat mast cells

Rat mast cells were challenged with compound 48/80 or calcium ionophore A23187, and protein kinase C activity in the cell pellets and the amount of histamine release into the supernatant were measured. After stopping the reaction, rat mast cells were lysed in a medium which prevents alterations in phosphorylation and dephosphorylation during sample processing. Histamine was significantly released from compound 48/80-stimulated mast cells at 30 s after the stimulation. In parallel with this, protein kinase C activity in the cell pellets increased at 30 s and 1 min and returned to basal value 3 min after the stimulation. When mast cells were incubated with various concentrations of 48/80 for 30 s, the amount of histamine release and protein kinase C activity increased dependently on the concentration of 48/80. Significant histamine release from A23187-stimulated mast cells was found at 1 min after the stimulation. Also protein kinase C activity in the cell pellets increased at 1 min and returned to basal value 5 min after the stimulation. A reduction of cytosolic protein kinase C activity was observed upon 48/80 treatment in a time- and concentration-dependent manner. Further, staurosporine, a potent inhibitor of protein kinase C, inhibited 48/80-induced histamine release in parallel with the inhibition of protein kinase C activity. These findings suggest that transient increase of protein kinase C activity may be involved in the mast cell activation process.     

Rapid induction of enterochromaffinlike cell tumors by histamine2-receptor blockade.

The effect of acid inhibition on gastric endocrine cells was investigated in Praomys (Mastomys) natalensis. Long-term treatment (1 to 32 weeks) with an irreversible histamine 2-receptor blocker (loxtidine) caused a sustained increase in plasma gastrin levels, which was accompanied by a gradual increase in histamine and histidine decarboxylase activity of the gastric oxyntic mucosa. The density of endocrine cells in the oxyntic mucosa increased gradually, doubled by 8 weeks, and was three times that of controls after 24 weeks of treatment. Hyperplastic changes in the endocrine cell population were evident after 2 to 8 weeks in all animals, whereas dysplastic or neoplastic lesions were observed in half the animals after 16, 24, and 32 weeks of treatment. Gross tumors in the oxyntic mucosa were observed in 1/4 of the animals treated for 24 or 32 weeks. Proliferating cells were identified as enterochromaffinlike cells because they were argyrophilic and immunopositive for chromogranin A and histamine. The results demonstrate that histamine 2-receptor blockade initiated by loxtidine promotes a rapid development of enterochromaffinlike cell tumors in Mastomys and suggest a critical role for gastrin in the formation of these tumors. However, the rate and frequency by which carcinoid tumors appeared in Mastomys after acid inhibition was much greater than that reported in other species, indicating that several factors, including hormonal and genetic factors, are important in the development of gastric endocrine tumors.     

Histamine and histidine decarboxylase: Immunomodulatory functions and regulatory mechanisms

Histamine is a bioactive monoamine that is synthesized by the enzymatic activity of histidine decarboxylase (HDC) in basophils, mast cells, gastric enterochromaffin‐like (ECL) cells and histaminergic neuronal cells. Upon a series of cellular stimuli, these cells release stored histamine, which elicits allergies, inflammation, and gastric acid secretion and regulates neuronal activity. Recent studies have shown that certain other types of myeloid lineage cells also produce histamine with HDC induction under various pathogenic stimuli. Histamine has been shown to play a series of pathophysiological roles by modulating immune and inflammatory responses in a number of disease conditions, whereas the mechanistic aspects underlying induced HDC expression remain elusive. In the present review, we summarize the current understanding of the regulatory mechanism of Hdc gene expression and the roles played by histamine in physiological contexts as well as pathogenic processes. We also introduce a newly developed histaminergic cell‐monitoring transgenic mouse line (Hdc‐BAC‐GFP) that serves as a valuable experimental tool to identify the source of histamine and dissect upstream regulatory signals.     

Histamine metabolism of the gastric mucosa following antrectomy

1. Histamine metabolism of the gastric mucosa in rats subjected to antrectomy, antrectomy and substitution with pentagastrin and exclusion of the antrum has been investigated employing various in vivo and in vitro methods.2. Mucosal histamine formation after antrectomy fell to about one third, whereas after antrum exclusion histamine formation increased many-fold.3. After antrectomy, mucosal histamine content decreased, and increased after antrum exclusion.4. After total gastrectomy, (a) whole-body histamine formation was reduced to about half, as judged by determining histamine excretion, and (b) pentagastrin infusion did not increase histamine excretion, showing absence of histamine mobilization from extra-gastric sources. In rats with the stomach retained, infusion of pentagastrin induced a dose-dependent increase in histamine excretion.5. Kinetic studies in which [(14)C]histidine was injected and the resulting urinary [(14)C]histamine determined showed that on pentagastrin infusion after antrectomy newly formed histamine was initially mobilized to a larger extent than in the controls.6. Antrectomized rats were subjected to substitution treatment by three injections per day of pentagastrin. After 3 weeks of substitution, histamine excretion was considerably higher than without substitution. After 6 weeks of substitution, histamine excretion was about the same in the substituted antrectomized, non-substituted antrectomized and sham-operated groups. Neither time nor substitution could, however, normalize the excretion of histamine on pentagastrin stimulation after antrectomy.7. In non-substituted antrectomized rats, pentagastrin was less effective in elevating mucosal histamine formation than in the substituted and sham-operated groups.8. The indispensability of the rat in this kind of study is emphasized.     

Histamine methyltransferase in dispersed cells from rabbit fundic mucosa

The cellular distribution of histamine N-methyltransferase was studied in rabbit gastric mucosa. The fundic mucosa was dispersed by collagenase treatment in Hanks' or calcium-free medium. In calcium-free medium, the number of dispersed cells/g wet tissue, as well as their viability was increased; histamine N-methyltransferase recovery was up to three-fold larger than in cells prepared in Hanks' medium. Furthermore, the calcium-free medium led to a greater acid secretory response, whereas the cellular pepsinogen content tended to be lower. Histamine N-methyltransferase activity was found in all cell fractions but was higher in the larger cell types. The enzyme activity showed only a partial correlation with either oxyntic or chief cells. These results indicate that the use of calcium-free medium to disperse and isolate rabbit mucosal cells improves cell quality. Histamine N-methyltransferase in the rabbit fundic mucosa, is found in more than one cell type, primarily the oxyntic and chief cells.     

Mast cell degranulation prior to ischemia decreases ischemia-reperfusion injury in the canine small intestine

OBJECTIVE: To determine the impact of previous mast cell degranulation on intestinal ischemia-reperfusion-induced mucosal damaged. MATERIALS: The hemodynamic and morphological consequences of complete arterial occlusion were evaluated in anesthetized dogs. The mast cell degranulator Cremophor-E1 (n = 5) and Compound 48/80 (n = 5) were used to investigate the involvement of gastrointestinal mast cells in ischemia-reperfusion-induced tissue reactions. Seven dogs subjected to complete segmental arterial occlusion served as controls. Intestinal biopsies taken at the end of 120-min ischemia and after 120 min of reperfusion were evaluated histologically. METHODS: The number of mast cells was determined and the degree of mucosal damage was evaluated according to the 5 grade Chiu scale. Mucosal histidine decarboxylase activity was measured in tissue samples and the rate of release of histamine was determined from the venous effluent of the ileal segment. RESULTS: In the control group, 120-min reperfusion significantly increased the plasma histamine level, and induced a severe tissue injury. In the compound 48/80 and Cremophor-E1-pretreated groups, the reduction in the baseline number of mast cells in the villi was 37% and 53%, respectively, and the ischemia-reperfusion-induced release of histamine was significantly decreased. In these groups, the basal mucosal histidine decarboxylase activity was significantly increased and the degree of damage of the intestinal mucosa was significantly reduced. CONCLUSION: It is proposed that mast cell degranulation prior to ischemia may induce a potentially protective mechanism in the small bowel mucosa and decreases ischemia-reperfusion injury in the dog.     

Effects of antrectomy or porta-caval shunting on the histamine-storing endocrine-like cells in oxyntic mucosa of rat stomach. A fluorescence histochemical, electron microscopic and chemical study.

1. The argyrophil (enterochromaffin-like) cells in the oxyntic gland area of the rat stomach contain histamine, which can be demonstrated fluorescence microscopically after exposure to gaseous OPT. After administration of L-dopa (or L-5-hydroxytryptophan), these cells produce and temporarily store dopamine (or 5-hydroxytryptamine), demonstrable by its characteristic formaldehyde-induced fluorescence. Ultrastructurally, the enterochromaffin-like cells, which have the appearance of polypeptide hormone-secreting cells, comprise two main cell types, the most predominant one having vesicular type granules (EGL cells), the second most predominant one having smaller, uniformly electron dense granules (A-like cells). 2. Rats were subjected to the following surgical treatments: antrectomy; porta-caval shunting; antrectomy+porta-caval shunting; or sham-operation. Three to eight weeks after surgery the histamine-storing cells (enterochromaffin-like cells) of the oxyntic mucosa were analysed by fluorescence histochemistry, light and (quantitative) electron microscopy, and fluorometric determination of amines. 3. After antrectomy, fluorescence histochemistry and silver staining revealed a reduced number of enterochromaffin-like cells. The histamine content in the oxyntic mucosa was reduced by about 50%. As in unoperated injection of pentagastrin seemed to mobilize histamine. Feeding or injection of insulin failed to do so in antrectomized as opposed to control rats. Ultrastructurally, the cytoplasmic granules of both endocrine-like cell types were less numerous than in the unoperated rats. The reduction in cell number and granularity was particularly conspicuous with regard to the EGL cells. 4. After porta-caval shunting the number of enterochromaffin-like cells increased markedly. Chemical determination revealed a twofold increase in the histamine concentration of the oxyntic mucosa. Feeding or injection of insulin or pentagastrin lowered the histamine concentration. As judged by electron microscopy, the proliferation of endocrine-like cells induced by porta-caval shunting was restricted to the ECL cell type. Besides occurring in greater number, these cells were larger than those in unoperated controls, and their cytoplasm was densely packed with granules that were increased in size. 5. Following antrectomy of the porta-caval shunted rats the number of enterochromaffin-like cells and the oxyntic histamine concentration was reduced. 6. The results support the idea that gastrin exerts trophic as well as excitatory effects on oxyntic endocrine-like cells.     

A modified schayer procedure for the estimation of histidine decarboxylase activity: Its application on tissue extracts from gastric mucosa of various mammals

The question of whether the gastric mucosa of mammals other than rats, mice and hamsters contains an acid (specific) histidine decarboxylase was re-investigated by using a modification of the classical isotope dilution method of Schayer. Its reliability was tested regarding sensitivity, specificity, precision and accuracy, applying criteria recommended by the International Federation of Clinical Chemistry. The assay is now suitable for measuring rather large series of samples.The suitability of several blanks for detecting histidine decarboxylase activity was investigated as a special problem of accuracy. The semicarbazide blank was found to be as reliable as the heating blank, the blank with tissue extract pretreated with perchloric acid or an incubation mixture without radio-labelled histidine.Reaction kinetics of histamine formation were linear over an incubation period of at least 3 h.Histidine decarboxylase activity at a rather low pH and substrate concentration, which is characteristic for the acid (specific) histidine decarboxylase, was demonstrated in the gastric mucosa of human subjects, dogs, rabbits and guinea-pigs, always using the same incubation conditions for all species investigated. The highest enzymic activity was present in the oxyntic mucosa, but could be measured also in other parts of the stomach.     

Histamine metabolism in lungs of rats infected withNippostrongylus brasiliensis

Several parameters connected to histamine metabolism and mast cell number were examined in the lungs of rats infected with the nematodeNippostrongylus brasiliensis. Histamine levels as well as mast cell numbers were found to be increased on day 14 after infection and were elevated during the whole time of the experiment. Histidine decarboxylase activity also reached a peak on day 14. There was no measurable activity of diamine oxidase in the lungs of parasitized and normal rats. It is postulated that the increase in histidine decarboxylase activity and histamine concentration observed in the present study is related to the process of mastocytosis.     

The role of tissue mast cells in exocrine secretion: Studies in the submandibular gland of the cat

Histamine was found to be stored in the submandibular gland of the cat mainly in the mast cells. The amine is released from mast cells by compound 48/80 and pilocarpine. This was demonstrated in thein vivo andin vitro experiments and by histological examination. During the physiological stimulation of the gland, via the electrical stimulation of the chorda tympani nerve, significant changes of histamine content were not found. This could be explained by the increased synthesis of histamine during physiological stimulation. The physiological role of histamine in salivary secretion was demonstrated. The similarity between the roles of histamine, mast cells and chromaffine-like cells in salivary and gastric secretion is discussed.     

Inhibition of histamine biosynthesis and gastric function in the rat: effect on pentagastrin-induced gastric acid secretion

A very simple and rapid method to measure gastric acid secretion in the conscious unoperated fasted rat is reported. Antisecretory activity of cimetidine and hypersecretory activity of pentagastrin are detected by this procedure. Pentagastrin induces gastric acid secretion and causes an increase by 2-fold in histidine decarboxylase activity. There is a delay between the increase of gastric acid output and the appearance of histidine decarboxylase activity. Co-administration of monofluoromethyl histidine, a specific irreversible inhibitor of histidine decarboxylase, does not modify the peak response to pentagastrin but shortens the duration of stimulation. We suggest from this data that histamine biosynthesis is needed for maintenance of elevated gastric acid secretion.     

Expression ofl -histidine decarboxylase (HDC) mRNA in rat stomach

We usedin situ hybridization with two 50-mer oligonucleotide probes, complementary to rat histidine decarboxylase (HDC) but with little or no sequence homology to rat dopa decarboxylase, to detect HDC mRNA in sections of rat stomach.A high number of silver grains indicating specific hybridization were detected on cells in the basal third of the gastric glands in the oxyntic mucosa with both probes suggesting that these cells express HDC similar to that of the fetal liver and adult brain. Submucosal mast cells were devoid of detectable hybridization. Our method enables quantitative studies on the regulation of the HDC gene in the rat stomach and other tissues.     

The effect of lowering the 5-hydroxytryptamine content of the rat spinal cord on analgesia produced by morphine

After prolonged fasting the activity of histidine decarboxylase in the oxyntic mucosa of the rat stomach is low. Feeding or injection of gastrin or insulin rapidly raises the enzyme activity. It was earlier suggested that all enzyme-activating agents act through release of gastrin. This view has found experimental support in studies which show that in antrectomized rats the enzyme is activated by gastrin but not by gastrin-releasing stimuli like feeding or vagal excitation (insulin hypoglycemia). In the present investigation rats were subjected to a variety of treatments and serum gastrin concentrations and gastric histidine decarboxylase activities were measured. The main findings were as follows.1. Feeding raised the serum gastrin level and the enzyme activity in unoperated rats. In fasted antrectomized rats the serum gastrin concentration was low; in freely fed antrectomized rats it was at the same level as in fasted unoperated rats. In antrectomized rats the enzyme activity was low and not raised by feeding.2. Acid in the antrum inhibits the release of gastrin whereas an alkaline pH may facilitate such release. All treatments that blocked acid secretion, thereby raising the antral pH, also raised the serum gastrin concentration and concomitantly the histidine decarboxylase activity. Thus, vagotomy increased the serum gastrin level and the histidine decarboxylase activity in fasted rats. Treatment of fasted unoperated rats with atropine or hexamethonium had similar effects. Antral exclusion, which prevents HCl from reaching the pyloric glands, resulted in marked increase in the serum gastrin concentration and in the enzyme activity of fasted rats.3. Injection of insulin resulted in a rather slow, progressive increase in the serum gastrin concentration. The peak was reached after about 4 hr. The enzyme activity was also raised markedly and the peak response occurred about 1 hr later.4. An increase in the histidine decarboxylase activity was invariably preceded or accompanied by a raised serum gastrin level. With fasted or fed unoperated, vagotomized, antrectomized or antrally excluded rats, the correlation coefficient for the relation between enzyme activity and serum gastrin concentration was 0.69 (P < 0.05).5. Porta-caval-shunted fasted rats responded to feeding or injection of insulin with marked activation of gastric histidine decarboxylase. The response after feeding was at least 5 times higher in shunted than in nonshunted rats but serum gastrin was only slightly higher. Following antrectomy of porta-caval-shunted rats feeding no longer raised the enzyme activity. Thus, the enzyme-activating agent was of antral origin. In the shunted rats injection of pentagastrin induced an enzyme activation about 5 times that seen in intact rats. This response was not significantly reduced by antrectomy.In conclusion, we have observed a correlation between serum gastrin concentration and histidine decarboxylase activity. We have failed to obtain evidence for the existence of any physiological intermediate other than gastrin in the activation of histidine decarboxylase induced by feeding, vagal stimulation or inhibition of acid secretion.     

Histamine content and synthesis in central and pheripheral nerve structures during stress

Histamine content and histidine decarboxylase activity in cortex and hypothalamus, together with histamine content in peripheral nervous structures were examined in normal and electrically stressed guinea-pigs. A significant increase in histidine decarboxylase activity in hypothalamus and cortex together with concomitant decrease in histamine content in hypothalamus have been found. Electric shock causes also a decrease in histamine content in spinal cord, spinal ganglia, dorsal roots and sensory nerve. The function of histamine in nervous system is discussed.     

Effect of gastric irradiation on gastric secretion and histamine in mice

Gastric irradiation selectively inhibits acid secretion. Histamine may have a role in the inhibitory process. Seven days after gastric irradiation with 9 Gy X-rays, mice underwent a stomach perfusion and secretion test. Under pentagastrin stimulation (62.5 g/kg), acid and histamine secretion was significantly lower than that of the unirradiated controls, while pepsin and potassium outputs were unchanged. Histamine concentration in the oxyntic region of the stomach assayed after the perfusion test was significantly lower than that of controls. It is postulated that the fall in the endogenous histamine store may contribute to the reduction in acid production.     

Prostaglandins and alkaline secretion from oxyntic, antral, and duodenal mucosa of the dog

Alkaline secretion (AS) measured under basal conditions in oxyntic and antral pouches of conscious dogs averaged about 20 mumol/30 min and was about three times lower than that from the duodenal pouch. Natural prostaglandin E2 and prostaglandin F2 alpha, but not prostaglandin I2, were effective stimulants of AS, mainly when given topically. Stable analogues such as 16,16-dimethyl prostaglandin E2 and prostaglandin I2 were relatively more potent stimulants than their parent prostaglandins (PGs), particularly when applied topically. The highest alkaline response of the oxyntic pouch to PG was about 5% of the maximal acid response of this pouch to histamine. Indomethacin reduced markedly AS from the duodenal but not from the oxyntic or antral pouch. AS from the duodenal pouch was relatively more sensitive than that from gastric pouches to the stimulation by PGs, which were effective also after pretreatment with indomethacin. This study shows that the oxyntic, antral, and duodenal mucosa of conscious dogs is capable of secreting bicarbonate, and this secretion, particularly from the duodenal mucosa, is highly sensitive to the stimulation with certain PGs, mainly of the E and F type and their analogues, and to suppression by indomethacin, a potent inhibitor of PG biosynthesis, suggesting that endogenous PGs are involved in the mechanism of AS.