Histamine Liberator Compound Research Articles

Determination of Histamine in Microdialysis Samples from Guinea Pig Skin by High-Performance Liquid Chromatography with Fluorescence Detection

Aim: To develop a sensitive and selective liquid-chromatographic method for the determination of histamine in microdialysis samples from guinea pig skin following allergenic provocation. Methods: The novel fluorescence derivatization method is based on an intramolecular excimer-forming reaction between 2 amino moieties of histamine and 2 molecules of 4-(1-pyrene)butanoyl chloride (PBC) yielding the corresponding dipyrene-labeled derivative. Results: The PBC derivative of histamine was separated within 20 min, and the detection limit (signal-to-noise ratio = 3) of histamine was 0.6 fmol/20 µl volume injected. The basal extracellular levels of histamine in guinea pig skin microdialysates were 20.6 ± 1.7 fmol/10 µl. Subcutaneous administration of histamine liberator compound 48/80 (3 mg/kg) increased the extracellular histamine levels in the skin dialysates by about 860%, whereas ovalbumin challenge (2 mg/kg i.v.) in the sensitized guinea pigs increased the extracellular histamine levels by about 3,030%. Conclusion: The novel technique for histamine determination in microdialysis samples from the guinea pig skin may be utilized in preclinical research of antihistaminergic drugs and evaluation of allergenic properties of various dermal preparations such as transdermal drug delivery systems.

Inhibition of complement dependent experimental inflammation in human skin by different heparin fractions

The anticoagulant activity of heparin is dependent on its affinity for antithrombin III (AT III) and on its molecular weight. In contrast, heparin fractions differing in these respects are equally effective inhibitors of the human complement system in vitro. In this study we designed and evaluated a model to investigate the effects of different heparin fractions on a complement dependent inflammation. Locally administered heparin, in a dose-dependent manner, inhibited the flare, itch and wheal responses induced by intradermal injection of heat-aggregated human IgG (HAGG). These reactions were also inhibited by the antihistamine mepyramine, favouring the view that HAGG activates complement and that the observed inflammatory response is mediated by anaphylatoxin liberation of histamine. Similar cutaneous reactions induced by trypsin, which can generate C3a and C5a by proteolysis of C3 and C5, the histamine liberator compound 48/80 or histamine were inhibited by mepyramine but not by heparin. Thus it is strongly suggested that heparin inhibits the HAGG induced reactions by modulating the early pre-C3 steps of complement activation. On a weight basis heparin fractions differing in AT III-affinity or in average molecular weight (5,000 and 16,000 D) were equally potent modulators of the HAGG-induced inflammation. We conclude that heparin can inhibit an apparently complement-dependent inflammation irrespective of its AT III-affinity or of its size, and suggest that a heparin with low anticoagulant activity could be of value as a modulator of inflammation and should be useful in investigating the consequences of complement inhibition in inflammation.

Mast cells and middle ear effusion

The mast cell--an important component of connective tissue--carries in its cytoplasmic granules various biologically active substances, such as heparin, histamine, and a broad spectrum of enzymes. This cell type plays a prominent role in inflammatory and allergic conditions. In the middle ear, the mast cells are mainly localized in the pars flaccida of the tympanic membrane and beneath the tracts of secretory and ciliated cells in the middle ear mucosa. Degranulation of the mast cells by the histamine liberator compound 48/80 causes histamine-rich effusion material to accumulate in the middle ear. Plugging of the eustachian tube and/or tympanic isthmus will bring about a similar accumulation. It would thus seem that mast cells in some way participate in the production of middle ear effusion, probably via their potent mediators.

Inhibition of the late phase reaction to anti-IgE by previous mast cell activation with compound 48/80.

The concept of an obligate association between mast cell activation and development of a late phase reaction (LPR) to various agents in human skin was further elucidated. Skin sites were treated four times at 24 h intervals with non-LPR-inducing doses of the histamine liberator compound 48/80 in 10 volunteers. The previously compound 48/80-challenged sites responded with an approximately 40% attenuated early response (P less than 0.01) and a 70% reduction of the LPR (P less than 0.001) to subsequently injected anti-IgE as compared with simultaneous reactions at control sites. The data suggest that mediators from the cutaneous mast cells are necessary for the final expression of an LPR.

Effect of neonatal treatment with capsaicin on carrageenan-induced paw oedema in the rat.

The time course of the paw oedema induced by the subplantar injection of carrageenan was studied in rats treated neonatally with capsaicin and in their vehicle-treated controls. In the capsaicin-treated rats, which show a permanent deficit of unmyelinated primary sensory neurones, carrageenan produced an oedema which was larger and lasted longer than in the vehicle-treated rats. Pretreatment with the histamine liberator compound 48/80 reduced the carrageenan-induced paw oedema only in the capsaicin-treated rats whereas pretreatment with indomethacin reduced it in both groups of rats. The increased and prolonged inflammatory response to carrageenan in capsaicin-treated rats may be explained by an enhanced release of histamine from mast cells and may also reflect a 'trophic disorder of the denervated skin'.

Histamine release from rat peritoneal mast cells exposed to ultraviolet light

After irradiation with ultraviolet light, rat peritoneal mast cells incubated with the histamine liberator compound 48/80 were found to have reduced capacity to release histamine. The action spectrum for histamine release inhibition seemed to be in the wavelength region of UVB. UV-light per se did not induce histamine liberation other than for dosages greater than or equal to 1.45 J/cm2. No such release was observed with the doses of UVA studied (3.4-20.5 J/cm2). The metabolic inhibitor 2.4-dinitrophenol (DNP), which almost completely inhibited compound 48/80-elicited histamine release, did not influence the UV-induced histamine release, indicating that the latter was not a secretory process but due to cytotoxic leakage of histamine from the cells. These results suggest that inhibition of histamine-releasing capacity or reduction of skin histamine due to photolysis of mast cells may explain the beneficial effect of UV in pruritic disorders where histamine release from mast cells is involved.

Influence of ultraviolet light on itch and flare reactions in human skin induced by histamine and the histamine liberator compound 48/80

The itch and flare responses induced by intradermal injection of histamine and the histamine liberator compound 48/80 were studied in healthy volunteers before and after exposure to UVB, UVA or PUVA administered 2-3 times weekly for 4 weeks. All three modalities were found to inhibit the responses induced by compound 48/80. The degree of tanning was most pronounced after PUVA and weakest after UVB, without any correlation between tanning and inhibition of itch. In contrast, when induced by histamine, the responses were not inhibited to the same extent, pronounced and significant inhibition being observed only for itching in subjects exposed to UVB. It is concluded that UVB, UVA and PUVA all might be of benefit in treating pruritic states if histamine release is involved and that UVB might have an additional effect by inducing hyposensitivity to itching stimuli.

Corticotropin/β-endorphin immunoreactivity in rat mast cells. Peptide or protease?

Rat mast cells were examined by immunohistochemistry and radio-immunoassay with antisera to various regions of the prohormone procorticottopin (ACTH)/β-endorphin. These antisera revealed specific immunostaining in mast cells obtained from the peritoneal and thoraic cavities and in sections of rat tongue, mesenterium, and lung. Addition of the histamine liberator compound 48/80 to suspensions of peritoneal-thoracic mast cells produced a large increase in the level of ACTH and β-endorphin in the medium as measured by radio-immunoassay. Although the released material in each case produced a displacement curve parallel to that of the known peptide, this phenomenon was attributed to degradation of labelled hormone by an apparent protease released from mast cells. ACTH immunoreactivity was not detected in cell extracts obtained with boiling water or acetic acid. These data strongly emphasize the potential artifacts that can occur when immunoreactivity alone is in the criterion for determining the presence of peptide hormones in cells.

Thermographic analysis of skin test reaction using AGA thermovision.

The course of infrared thermography including isothermograms on the skin surface was investigated considering blood flow, redness of the skin and permeability of blood vessel, in the following skin reactions: 1. Intracutaneous injection of histamine and histamine liberator compound 48/80 increased the heat radiation. Local application of antihistamine externa which decreased the development of the urticarial histamine reaction, increased the infrared radiation of the skin surface. Combined injection of histamine or histamine liberators with antihistamines in a sufficient dosis (1:1 respectively 4:1) diminished also the heat radiation in addition to the urticarial reaction. 2. The Pyrexal reaction of the skin with early erythema and later papule development shows an equivalent picture in the AGA Thermovision. The pretreatment shows an equivalent picture in the AGA Thermovision. The pretreatment of the skin with corticosteroid ointments shows a corresponding lowering of the erythema, of papule development as well as of heat radiation. The blanching of corticosteroids after occlusive dressing is difficult to recognize by the isotherms of AGA Thermovision. 3. Allergic reactions of the immediate type show, corresponding to the wheal eruption, a marked increased of heat radiation combined with a projection of the enlarged veins on the skin surface. 4. Allergic reactions of the delayed type are combined with a definite elevation of heat radiation of the skin. The area of a positive skin test with allergic eczematous reaction shows a distinct elevation of ann infrared radiation. Although the allergic skin area which was substantiated by a positive skin test was no longer visible, a distinct infrared radiation could be detected. Preventive treatment of the test area of skin patch-testing with corticosteroids inhibits the heat radiation even if the allergic eczematous reaction occurs faintly. The thermographic analysis of the different skin test reactions complied with the morphological aspects of the reaction.

Effect of Terbutaline, a β2‐adrenergic Receptor Stimulating Compound, on Cutaneous Responses to Histamine, Allergen, Compound 48/80, and Trypsin

The beta-adrenoceptor stimulating agent terbutaline (2 ng-2 microgram) injected intradermally in eight atopic subjects produced a dose-dependent inhibition of the skin reactions induced by subsequently injected allergen. After injection of 0.5 microgram terbutaline inhibition of the flare and weal responses was demonstrable throughout the observation period of 90 min. The flare response induced by histamine, the histamine liberator compound 48/80 and the proteolytic enzyme trypsin was not inhibited by terbutaline in the doses used, suggesting a selective action of terbutaline on the allergen-induced response. The weal response elicited by histamine and compound 48/80 was slightly reduced by 2 microgram terbutaline. It is suggested that pretreatment of the skin with terbutaline interferes with the ability of the cutaneous mast cells to respond to challenge with allergen and that terbutaline produces this effect in doses lower than those needed to counteract the permeability increasing effect of released mediator substances.

The effect of prostaglandin on iridial blood vessel permeability.

Light and electron microscopy is used to examine the effect of exogenous PGE1 on the permeability and reactivity of rat iridial blood vessels. Results show that topical PGE1 causes an increase in the permeability of iridial vessles to carbon particles (200 A diameter). The technique of carbon labelling is used to quantitate increases in permeability caused by varying concentrations of PGE1 (0.001-1.0 mg/ml). Regression analysis shows that there is a linear relationship (P less than 0.02) between carbon labelling and PGE1 concentration over the range of concentrations tested. In other experiments rats were treated with the systemic histamine liberator Compound 48/80, or with topical applications of histamine diphosphate in order to examine the effects of exogenous and endogenous histamine upon iridial blood vessel permeability. These procedures produce only minimal labelling of iridial vessels. It therefore seems likely that PGE1 has a direct effect on iridial vessels and does not act indirectly by bringing about the liberation of endogenous histamine.

In vitro effects of histamine liberator compound 48/80 on rat superior cervical ganglia with special regard to the small granule-containing cells

The effect of the histamine liberator compound 48/80 on the rat superior cervical ganglia in vitro has been investigated. After incubation of the ganglia with compound 48/80: (1) ganglionic mast cells degranulate in the same manner as in other tissues; (2) cell bodies of the postganglionic neurons are not affected by compound 48/80; (3) there is evidence that ganglionic interneurons, the monoamine-containing cells are activated. The ultrastructural aspects of this process are characterized by degranulation of perikarya and accumulation of dense core vesicles in cell processes and in terminals adjacent to presynaptic membranes. These vesicles vary in size between 200-800 A in diameter. They may represent storage sites of the neuro-transmitter complexes that have undergone exocytosis. The results are discussed with special reference to models of exocytotic processes involving the adrenergic transmitter. It is concluded that monoamine-containing cells represent interneurons in sympathetic ganglia which inhibit ganglionic transmission and which are stimulated by low concentrations of compound 48/80 in vitro.

Studies on the mechanism of decidualization. III. Decicualization in the histamine-depleted rat.

Pregnant and pseudopregnant rats were depleted of hista¬ mine by treatment with histamine liberator Compound 48/80 or pyrathiazine. The response to decidual induction by systemic histamine release was compared with the response to direct stimulation of the endometrium. The decidual reaction in response to endometrial trauma and to the presence of ova was markedly reduced, whereas systemic induction with pyrathiazine could be prevented completely. These observations support the hypothesis that induction of decidualization by systemic pyrathiazine is mediated by the release ofhistamine systemically, whereas induction by local stimuli is mediated by release of uterine histamine only.

Mast cell degranulation in the rat uvea induced by histamine-liberator compound 48/80.

Compound 48/80, a condensation product of<i>p</i>-methoxyphenethyl-methylamine and formaldehyde, has been shown to cause the liberation of histamine from tissue mast cells of the rat and certain other species.^3,9,10,12Liberation of the histamine is accompanied by degranulation of the mast cells. Compound 48/80 has, however, been shown to be less effective as a histamine liberator in certain other species, in particular the guinea pig.²The presence of mast cells in normal animal eyes was reported by Jorpes, Holmgren, and Wilander in 1937,⁷and changes in the mast cell distribution in certain clinical conditions in human eyes have been reported by several investigators.^6,13,14Further, in recent studies, Larsen has shown that there are cytological changes in the mast cells of the uveas of experimental animals after experimentally induced uveitis.⁸ The object of this study was to determine whether the mast cells localized in the eyes

EFFECT OF MAGNESIUM DEFICIENCY ON THE REGENERATION OF MAST CELLS AFTER TREATMENT WITH 48/80 IN RATS

The decrease in subcutaneous mast cell population observed in rats on a magnesium-deficient diet has been confirmed. Pretreatment with histamine liberator compound 48/80 accentuates the decrease in mast cell counts. This condition was apparently the result of defective regeneration. The decrease in the number of subcutaneous mast cells was accompanied by a parallel reduction of histamine as appreciated by the guinea pig ileum assay. The peripheral vasodilation, which is an early sign of magnesium deficiency, did not appear in rats pretreated with 48/80.

EFFECT OF MAGNESIUM DEFICIENCY ON THE REGENERATION OF MAST CELLS AFTER TREATMENT WITH 48/80 IN RATS

The decrease in subcutaneous mast cell population observed in rats on a magnesium-deficient diet has been confirmed. Pretreatment with histamine liberator compound 48/80 accentuates the decrease in mast cell counts. This condition was apparently the result of defective regeneration. The decrease in the number of subcutaneous mast cells was accompanied by a parallel reduction of histamine as appreciated by the guinea pig ileum assay. The peripheral vasodilation, which is an early sign of magnesium deficiency, did not appear in rats pretreated with 48/80.

HISTAMINE RELEASE BY HOMOLOGOUS PLASMA IN THE DOG

Dogs were injected intracutaneously with their own plasma and with plasma from other dogs. The injected skin in each case was excised and incubated in Tyrode's medium. More free histamine was found in the media containing skin injected with homologous plasma than in the media containing skin injected with autologous plasma. When considered with earlier observations, this result justifies the conclusion that, in the dog, homologous plasma releases histamine on initial injection. Similar experiments with the histamine liberator compound 48/80 showed the method to be extremely sensitive in detecting histamine release.

HISTAMINE RELEASE BY HOMOLOGOUS PLASMA IN THE DOG

Dogs were injected intracutaneously with their own plasma and with plasma from other dogs. The injected skin in each case was excised and incubated in Tyrode's medium. More free histamine was found in the media containing skin injected with homologous plasma than in the media containing skin injected with autologous plasma. When considered with earlier observations, this result justifies the conclusion that, in the dog, homologous plasma releases histamine on initial injection. Similar experiments with the histamine liberator compound 48/80 showed the method to be extremely sensitive in detecting histamine release.

Release of histamine by the histamine liberator compound 48/80 in cats.

The Journal of PhysiologyVolume 121, Issue 3 p. 517-538 ArticleFree Access Release of histamine by the histamine liberator compound 48/80 in cats A. N. Smith, A. N. SmithSearch for more papers by this author A. N. Smith, A. N. SmithSearch for more papers by this author First published: 28 September 1953 https://doi.org/10.1113/jphysiol.1953.sp004962Citations: 22AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat Citing Literature Volume121, Issue3September 28, 1953Pages 517-538 RelatedInformation