W. Schmutzler

Histaminasefreisetzung durch heparin und protamin beim meerschweinchen

Abstract Injections of heparin have been shown to provoke very strong histaminase activity in the plasma of guinea pigs, but not in other species so far tested (rat, rabbit, dog, hen, man). The activity is much stronger than in anaphylactic shock, when more than 50 IU/kg of heparin is given. Like the anaphylactic histaminase the heparin induced plasma histaminase comes from the liver. This has been demonstrated by injections of heparin in different sections of the cardiovascular system as well as by studies of the effect of heparin on the heart-lung-preparation and heart-lung-liver-preparation. The histaminase activity of the plasma is completely abolished by aminoguanidine. Protamine (as sulphate), too, has a strong histaminase liberating effect on the liver, 5 mg being as effective as 50 IU heparin. Protamine and heparin, on the other hand, neutralize each other when given simultaneously by two different intravenous routes. In this case the dose relationship is 1 mg protamine sulphate to 50 IU heparin.

ber die Wirkung von Heparin und Aminoguanidin auf den anaphylaktischen, Anaphylatoxin-und Histamin-Schock des Meerschweinchens

SummaryThe discovery, previously communicated, that heparin is a very strong histaminase liberator in guinea pigs incited to reinvestigate on a greater basis the unsolved problem of heparin action on anaphylactic shock as well as on the anaphylatoxin-(AT-) and histamine shock.To establish that heparin effects observed were due to histaminase release aminoguanidine was given together with heparin in special experiments.Aminoguanidine alone was also given in some experiments to detect any influences on shock of that part of histaminase which is released into the blood during anaphylactic as well as (to a lesser degree) AT-shock (but not histamine shock) by the shock itself.Shock rate and death rate were registered and plasma histamine level measured by taking blood from the carotid artery 1 1/2 min after the challenging injection (into the V. jugul.). The heparin dose was 500 IU/kg and the aminoguanidine (as sulphate) dose 2 or 10 mg/kg.The following results were obtained:1.There was no significant influence on anaphylactic shock of heparin or aminoguanidine when using ovalbumin as antigen in doses of 1, 2 or 10 mg/kg. The plasma histamine level was significantly decreased by heparin only in the experiments with 1 mg ovalbumin/kg.2.In histamine shock (provoked by injection of 0.36 mg histamine (as base)/kg) heparin reduced the shock rate and death rate as well as the histamine level significantly and was ineffective against 0.6 mg histamine/kg. The heparin effects were completely prevented by aminoguanidine (2 mg/kg.)3.Against the shock provoked by two different doses of AT (dextran treated rat serum) heparin was effective by lowering the shock and/or death rate in most instances and histamine level in a significant manner. These effects could be reduced or prevented by aminoguanidine pretreatment.4.Aminoguanidine alone increased the shock and death rate only in histamine shock (by 0.36 mg/kg) depending on the aminoguanidine dose. There was no influence at all on histamine level to be seen at the time of withdrawing blood.

Untersuchungen ber Substanzen mit histaminaseliberierender Wirkung beim Meerschweinchen

SummaryProsecuting our investigations on the histaminase-liberating action on the guinea pig liver of heparin and protamine we studied the effects of several other related compounds by injecting into the portal vein and estimating the histaminase activity in the hepatic vein plasma.Besides heparin following polyanions are very effective histaminase-liberators: Pentosanpolysulphate, polyethylenesulphonate, degraded carrageenan, polyvinylsulphate and dextransulphate. There is no significant difference between the action of dextransulphate of low and heigh molecular weight. Agar which contains only few ester sulphate groups liberates clearly less histaminase. Non-sulphated dextrans, the non-polymerised dodecylsulphate and sodium sulphate are ineffective.The histaminase-liberating action of the polycation clupein is comparable to salmin. It is more effective than agar, but less than the other polyanions. Spermine and 1-arginine were ineffective.A weak but significant release of histaminase was observed after intraportal injection of trypsin or EDTA and citrate, whereas calcium-EDTA was ineffective.

Über die Herkunft der Plasmahistaminase im anaphylaktischen Schock des Meerschweinchens

SummaryHistaminase is released into the blood during anaphylactic shock in guinea-pigs. Experiments are reported in which the antigen is applied to different sections of the cardiovascular system and to the heart lung preparation. These experiments demonstrate that the liver is the site of histaminase release in guinea-pigs.

Histamine-Independent Mechanisms in Shock in Guinea-Pigs

Author’s address: Prof. Dr. H. Giertz, Pharmacological Institute of the University, Katharinenstrasse 29, 78 Freiburg i. Br. (Germany) Our investigations as to the role of histamine in anaphylactic, anaphylatoxin and Forssman shock detected some mechanisms which are not induced by the release of histamine. For example, in contrast to anaphylatoxin shock, large doses of antihistamines into saline perfused guinea-pig lungs did not influence the intensity either of the anaphylactic bronchospasm or of the constriction of lung vessels. In the blood-perfused heart-lung preparation of guinea-pigs, the anaphylactic histamine release was as high as in the isolated lung, and the anaphylactic bronchoconstriction was not inhibited by antihistamines. In most preparations, the cardiac output and the blood pressure did not change until heart failure induced by anoxia was apparent. In some experiments heart failure was observed immediately after antigen injection. This early heart failure occurred more frequently in the presence of antihistamines, suggesting that an initial small anaphylactic damage to the heart may have been made visible, enlarged and aggravated by the antihistamine. The early heart failure may be the result of anaphylactic constriction of the pulmonary vessels. In the heart-lung-liver preparation of guinea-pigs, anaphylactic constriction of the liver vessels probably occurred and resulted in decreasing cardiac output, diminished heart volume and depressed blood pressure in the right atrium. In the intact animal, anaphylatoxin shock produced a decrease in the number of white blood cells but this decrease has nothing to do with the bronchospasm as this occurred when red blood cell G i e r t z et al. 47 suspensions were perfused in the heart-lung preparation. Anaphyla-toxin as well as anaphylaxis had an antihistamine-resistant effect on the liver vessels. The acute mortality of guinea-pigs due to anaphylactic bron-chospasm was totally inhibited by antihistamines, but many of the animals died later with congestion of the abdominal organs and dilatation of the right ventricle, but no inflation of the lungs. Perhaps this form of protracted shock may be induced by the antihistamine-resistant constriction of the pulmonary and liver vessels. Somewhat larger doses of antihistamines were needed to reduce the acute mortality in anaphylactic shock than in anaphyla-toxin shock but this may result from the fact that histamine released from the lung itself plays a major part in anaphylaxis in this species.

Histaminase in Shock in Guinea-Pigs

Author’s address: Prof. Dr. F. Hahn, Dept. of Pharmacology, University of Freiburg, 78 Freiburg i. Br. (Germany) During the anaphylactic shock in guinea-pigs a very strong histaminase activity appears in the plasma. This is not the case in Forssman shock, whereas in the anaphylatoxin shock also a histaminase activity occurs, but of lower strength. Experiments showed that the liver is the main source of the histaminase. Antigen injected into the portal vein of sensitized guinea-pigs resulted in a strong histaminase activity occurring in the hepatic vein and this activity was inhibited by aminoguanidine. Since the histaminase appeared very quickly after the antigen injection we assumed that it is released from some sources in the liver. A constituent of liver cells other than histamine might be involved. It was soon found that heparin produced maximal release of histaminase and that the heparin-induced histaminase originated in the liver like in anaphylactic shock. There was no increased activity of plasma histaminase in a heart-lung preparation perfused with heparin-containing blood but as soon as the liver was included in the circulation activity occurred. Of interest was the finding that protamine sulphate (an antagonist of heparin) also produced a strong increase in histaminase activity in the plasma of the intact guinea-pig. 5 mg protamine sulphate have the same effect as 50 units of heparin. However, heparin and protamine inhibited each other in these activations, 1 mg protamine neutralizing 50 units of heparin. Histaminase release may perhaps take place when heparin-like substances react with certain basic proteins in tissues, particularly liver tissue.

Zur Frage der Heparin-Beteiligung bei der anaphylaktischen Histaminase-Freisetzung des Meerschweinchens

Die Blepharoptosis is~ eine charakteristische Wirkung yon l%eserpin, die zum biologischen Nachweis reserpinartig ~4rkender Verbindungen benutzt wird (Rv~IN u. Mitarb., 1957). Die Verengung der Lidspalte kommt dadurch zus~ande, dab der sympathisch innervierte, glattmuskelige Antefl des ]V[. levator palpebrae ausfi~llt. Diese glatte Musku]atur (= Mtillerscher Muskel des Lides) ist f/it die tonische Funk~ion der LidspaltenSffnung verantwortlich. Im allgemeinen wird die Ptosis auf einen zentralen Effekt yon Reserpia zur/ickgeffihrt. Andererseits interferiert die Noradrenalin-Entleerung nach Reserpin auch mit dem peripheren adrenergen Ubertragungsmechanismus an der sympathisch innervierten glatten Muskulatur. Durch Ableitung der elektrischen Akti~d~/~t im zuffihrenden postganglion~ren Sympathicus-Ast sollte gepriift werden, ob eine mit dem Auftreten der Ptosis zeitlich korrelierte Abnahme der Impu]st~tigkeit zu beobachten ist. An flach narkotisierten und ~eflcurarisierten Katzen wurden Einzelfasern und Filamente des zum Mfillerschen Muskel ziehenden postganglion~ren Sympathicusastes hergestellt. Nach i.v. Injektion yon 1,0 mg/kg Reserpin kam es nach einer initialen Zunahme der Aktivit~ (bis 30 rain) zu einer mgBigen Einschrgnkung der Entladungst~tigkeit (nach 2 Std), die aber nicht ausreicht, die auftretende Ptosis zu erkli~ren. Es handelt sich bei der Reserpinptosis nach diesen Versuchen in erster Linie um ein peripher ausgelSstes Phgnomen. Auch die antagonistische Wirkung yon Cocain gegen/iber der Reserpinptosis scheint auf einem peripheren Angriffspunkt zu beruhen. Literatur Ru~IN, B., M. H. MM, o~, ~. H. WAvo~, and J. C. Bv~K~: J. Pharmaco]. exp. Ther. 120, 125--136 (1957). Priv.-Doz. Dr. G:ERttARD SCHlV//DT, Pharmakologisches Institut der Universit~t, 3400 G6ttingen, Geis~str. 9

Hemmung der anaphylaktischen histaminasefreisetzung durch protamin beim meerschweinchen

Abstract Previous experiments had shown that in guinea pigs the liver is the source of the plasma histaminase during the anaphylactic shock as well as after injections of heparin. Furthermore, the liver of guinea pigs is also involved in the anaphylactic release of heparin-like substances which have a histaminase releasing activity. The present study indicates that the release of anaphylactic histaminase into the plasma is mediated by heparin. In sensitized guinea pigs the plasma histaminase activity after intravenous injection of antigen (1 mg ovalbumin/kg) is strongly reduced by simultaneous injection of protamine (1 or 2 mg/kg) into a different vein, although higher doses (5 mg/kg or more) of protamine have like heparin a strong histaminase releasing effect. The effect of protamine on the anaphylactic histaminase release is in accordance with its effect on the histaminase release produced by heparin injection. Heparin doses which provoke the same plasma histaminase activity as the anaphylactic shock are also neutralized by 1 mg protamine/kg. In vitro , protamine has no inhibiting action on the activity of plasma histaminase. Given in different doses, heparin revealed no inhibitory effect on the anaphylactic histaminase release. Protamine is ineffective against the anaphylactic histamine release into the plasma of guinea pigs and does not alter the acute or subacute death rate in shock.

Darstellung eines in vitro wirksamen, Histamin und acetylcholin sensibillisierenden Faktors der Haemophilus Pertussis-Vaccine

~) PFEIF~ER, H.H.: Naturwissenschaftea 46, 12 (1959). =)RmNER, M.: In Handbueh der Physik, Bd. 6, S. 434, 535. BerlinG6ttiilgen-Heidelberg: Springer t958. Vgl. aueh die Diskussion der Differenz zwischen eIastisehen Solen und relaxierenden Gelen bei REINER, M.: Bull. Res. Council Israel 1, 5 (195~). -a) PFEIFFER, B.H.: Das Polarisationsmikroskop als Meginstrument in Biologie und Medizin, S. 55, 64. Braunsehweig: Friedr. Vieweg & Sohn 1949. ~) SWANN, iV[.l~., II. tiM. MITCHISON: J. Exp. Biol. 27, 226 (~950). ~)NITSeHg~NN, H., u. J. SCHRAOE: Hetv. chim. Acta 31, 297 (1948). e) SCOTT BLAIR, O.W.: Survey of general and applied theology, p. 159. London: Pergamon Press t949. REINER, .IV[. : S. ~), S. 451.

Untersuchungen zur substrat- und inhibitor-spezifität der freisetzbaren meerschweinchenleberhistaminase☆

Abstract The substrate- and inhibitor specificity of the histamine metabolizing enzyme released from the guinea pig liver by heparin has been studied. During the degradation of histamine 1 2 mole oxygen per mole substrate is consumed. The reaction velocity does not depend on the substrate concentration in the range from 4.5 × 10 −6 to 1 × 10 −2 M. Besides histamine the diamines cadaverine and putrescine, the monoamines β-phenylethylamine and benzylamine, and the polyamine spermidine are oxidized. A significant degradation of 5-hydroxytryptamine, tryptamine, mescaline, and tyramine could not be detected by the Warburg-technique. 4-aminobutyraldehyde (as Δ 1 -pyrroline) and benzaldehyde were demonstrated to be reaction products of the degradation of putrescine and benzylamine respectively. The pH-optimum of the degradation of cadaverine is 6.0, of histamine 7.0. and of benzylamine 8.0. A complete inhibition of the degradation of histamine, cadaverine, and benzylamine was achieved with aminoguanidine (10 −6 M), hydroxylamine (10 −5 M), or semicarbazide (10 −4 M). An incomplete inhibition was observed with isonicotinic acid hydrazide (10 −2 M) and iproniazide (10 −2 M). No inhibition at all was observed with 2-phenylcyclopropylamine (10 −3 M). It is concluded that the enzyme belongs to the carbonyl-reagent sensitive amine oxidases (Enzyme Commission No. 1.4.3.6).