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Mechanisms of histamine release from rat mast cells induced by polyethylenimines and polyallylamines
710
Our results show that thymopentin is a weak histamine releasing peptide. The release of amine from rat mast cells by thymopentin has characteristics of histamine release caused by other polyamines (Erjavec et al., 1981; Devillier et al., 1985).
erences Deviilier. P.. Renoux. M.. Giroud. J.P.. Regoli, D.. 117. Eur. J. Pharmacol., 89. Erjavec. F., Lembeck. F., Irman-Flojanc. T.. Skofitsch, G., Donr.erer, J., Saria. A., Holzer,P., 1981. P. Naunyn-Schiedeb. Arch. Pharmacol. 317.67. Malaise, M-G., Hazee-Hagelstein, M.T.. Reuter, A.M., Vrinds-Gevart. Y., Goldstein, G.C.. Franchimont, P., 1987, In: Immune Regulation by Characterized Polypeptides. Allan R. Liss. Inc., 111.
! I
P.tu.011
so
Yoshino,
ylenimines
ist
Y., Nagaya,
K., Sekino,
H., U&da,
M.K. and Suzuki-Nishimura,
Department of Molecular Pharmacoiogv, Me@ College of Pharmacy, 1-35-23 Nozawa, Setagaya-ku,
T.
Tokyo 154, Japan
Our group previously reported that synthetic polycations with various molecular weights [polyethylenimines @Is) and polyallyiamines (PAAs)] released histamine from rat mast cells tguzuki-Nishimura et al., 1989). These polymers are essentially harmless to humans and are used as additives in various papers. PEIs and PAAs are simpler than compound 48/80 in chemical structure and are stable in aqueous solution. PEIs and PAAs (l-10 pg/ml) released histamine from rat peritoneal mast cells in the presence of calcium. Acetylated derivatives and triethylentetramine did not induce histamine release. Unlike their fusogenic activites (Oku et al., 1986), histamine-releasing potencies of these polymers did not depend on their molecular weights. We, therefore, compared the histamine release induced by PEta and PAAs with that induced by compound 48/80. The histamine release induced by compound 48/80 was known to be inhibited by lidocaine and islet activating protein, but not by disodium cromoglycate. The histamine releases induced by all PEIs and PAAs tested were inhibited by lidocaine (1 mM), but not by disodium cromoglycate (3 PM). Islet activating protein inhibited the histamine release induced by PEI with a molecular weight of 600 (PEI,). Its effects on the release by other PEIs and PAAs were less than that on PEI,. It is likely that the inhibition of G proteins by the islet activating protein resulted in a decrease in the histamine release. This possibility was supported by the finding that guanyl-5’-( /?, y-imino) triphosphate enhanced the histamine release. PEI, was further studied because the effect of PEI, seemed to resemble that of compound 48/80. We compared the binding sites for PEI, with those for compound 48/80 using various lectins. Under our conditions, concanavalin A (Con A), phytohemagglutinin (PHA) and wheat germ agglutinin (WGA) alone were unable to induce the histamine release, because of the absence of phosphatidyl-serine. The release induced by 3 rg/ml PEI, was inhibited by WGA and PHA, but not by Con A. The binding of PEI, to mast cells seemed to be inhibited by WGA rather due to its affinity for sialic acid than to its affinity for GluNAc, because Limax flavus agglutinin (LFA), a specific lectin for sialic acid, also decreased the release of histamine induced by PEI, (3 pg/ml) from 41.1 + 2.0 (in the absence of LFA. n = 6) to 13.5 f 3.0 (in the presence of 100 pg/ml LFA, n = 6) of the total cell content. Moreover, the release of histamine induced by compound 48/80 was inhibited by WGA and PHA. but not by Con A, suggesting that the binding sites for PEI, and compound 48/80 had siak acid and GalNAc residues. The binding sites for PEI, seemed to especially overlap those of compound 48/80. Therefore, the binding of PEI, to their binding sites of mast cells seemed to activate G protein to release histamine from the mast cells. It is possible that mast cells also have other binding sites for PAAs, because WGA, PHA and Con A partially inhibited the histamine release induced by PAA with a molecular weight of 10,000. Thus PEIs and PAAs should be useful in further studies on the mechanisms of signal transduction coupling with G proteins of rat mast ce!!s.
711
References Suzu~-IWGmw T., SefiW H..Yoshind,Y., Nagaya, K.. Oku, N.. Nango, M. and Uchida, M.K., 1989, Jpn. N., Yamaguchi, N., Yamaguchi, N., Shibamoto, S., Ito. F. and Nango. M., 1986, J. B&hem. It-~o, 935.
J_ Ph~acol.51.279_
Oh
El
P.tu.012
relaxing factor an
a!st ee
i&amine release Mannaioni,
P.F., Masini, E., Salvemini
*, D., Pistelli, A., Korbut *, R. and Vane *, J.R.
Dept. oj Preciinical and Clinical Pharmacology, Viale G. B. Morgagni 65, 50134 Florence. Italy and * The WiU:am Harvey Research Institute, St. Bartholomew’s Hospital Medical College, Charterhouse Square. London EC JM 6 BQ, U.K.
Activated endothelial cells release a labile factor which relaxes vascular smooth muscle and inhibits platelet aggregation. This endothelium-derived relaxing factor (EDRF) is accounted for by nitric oxide (NO) synthetised from the amino-acid I-arginine, acting via stimulation of soluble guanylate cyclase and whose biological actions are terminated by superoxide anion and by oxy-haemoglobin (Moncada et al., 1989). Beside endothelial cells, relaxing factors have been shown to be released by activated neutrophils. macrophages. hepatocytes, Kupfer cells and neurons (Griffith et al., 1989). Here we show that isolated rat serosal mast cells ‘release a factor that inhibits p!atelet aggregation and has a pharmacological profile similar to that described for EDRF (NO). In turn, drugs which are capable of generaring NO inhibit mast cell histamine release induced by different stimuli. Incubation of mast cells with washed human platelets resulted in an inhibition of thrombine-induced platelet aggregation that was dependent on the number of the cells added. The inhibition was potentiated by superoxide dismutase and reversed by oxy-haemoglobin. Mast cells are more potent than bovine aortic endothelial cells in their ability to release a nitric oxide-like factor. A NO-generating drug (sodium nitroprusside. NaNP) was shown capable of inhibiting the release of histamine induced by compound 48/80, Ca2+ ionophore A 23187 and by a free radical-generating system (paracetamol+ prostaglandin-H-synthetase, PHS) in rat serosal mast cells. The inhibiting action was concentration-dependent. had a long time-course, was potentiated by the presence of an endotoxin lipopolysaccharide and was reversed by OXYhaemoglobin. These data show that the nitric oxide-like factor modulates the release of mast cell histamine. The work carried out at the William Harvey Res. Institute was supported by a grant from Glaxo Group Research Ltd. References Moncada, S., Palmer, R.M.J. and Higgs. E.A., 1989.
Biosynthesis
of nitric
oxide
from
L-Arginine.
1709-1715. Griffith,
T. and Randall,
M.. 1989. Nitric
oxide comes of age. The Lancet. October 7. 875-876.
Biochem.
Pharmacol.
38.
Our results show that thymopentin is a weak histamine releasing peptide. The release of amine from rat mast cells by thymopentin has characteristics of histamine release caused by other polyamines (Erjavec et al., 1981; Devillier et al., 1985).
erences Deviilier. P.. Renoux. M.. Giroud. J.P.. Regoli, D.. 117. Eur. J. Pharmacol., 89. Erjavec. F., Lembeck. F., Irman-Flojanc. T.. Skofitsch, G., Donr.erer, J., Saria. A., Holzer,P., 1981. P. Naunyn-Schiedeb. Arch. Pharmacol. 317.67. Malaise, M-G., Hazee-Hagelstein, M.T.. Reuter, A.M., Vrinds-Gevart. Y., Goldstein, G.C.. Franchimont, P., 1987, In: Immune Regulation by Characterized Polypeptides. Allan R. Liss. Inc., 111.
! I
P.tu.011
so
Yoshino,
ylenimines
ist
Y., Nagaya,
K., Sekino,
H., U&da,
M.K. and Suzuki-Nishimura,
Department of Molecular Pharmacoiogv, Me@ College of Pharmacy, 1-35-23 Nozawa, Setagaya-ku,
T.
Tokyo 154, Japan
Our group previously reported that synthetic polycations with various molecular weights [polyethylenimines @Is) and polyallyiamines (PAAs)] released histamine from rat mast cells tguzuki-Nishimura et al., 1989). These polymers are essentially harmless to humans and are used as additives in various papers. PEIs and PAAs are simpler than compound 48/80 in chemical structure and are stable in aqueous solution. PEIs and PAAs (l-10 pg/ml) released histamine from rat peritoneal mast cells in the presence of calcium. Acetylated derivatives and triethylentetramine did not induce histamine release. Unlike their fusogenic activites (Oku et al., 1986), histamine-releasing potencies of these polymers did not depend on their molecular weights. We, therefore, compared the histamine release induced by PEta and PAAs with that induced by compound 48/80. The histamine release induced by compound 48/80 was known to be inhibited by lidocaine and islet activating protein, but not by disodium cromoglycate. The histamine releases induced by all PEIs and PAAs tested were inhibited by lidocaine (1 mM), but not by disodium cromoglycate (3 PM). Islet activating protein inhibited the histamine release induced by PEI with a molecular weight of 600 (PEI,). Its effects on the release by other PEIs and PAAs were less than that on PEI,. It is likely that the inhibition of G proteins by the islet activating protein resulted in a decrease in the histamine release. This possibility was supported by the finding that guanyl-5’-( /?, y-imino) triphosphate enhanced the histamine release. PEI, was further studied because the effect of PEI, seemed to resemble that of compound 48/80. We compared the binding sites for PEI, with those for compound 48/80 using various lectins. Under our conditions, concanavalin A (Con A), phytohemagglutinin (PHA) and wheat germ agglutinin (WGA) alone were unable to induce the histamine release, because of the absence of phosphatidyl-serine. The release induced by 3 rg/ml PEI, was inhibited by WGA and PHA, but not by Con A. The binding of PEI, to mast cells seemed to be inhibited by WGA rather due to its affinity for sialic acid than to its affinity for GluNAc, because Limax flavus agglutinin (LFA), a specific lectin for sialic acid, also decreased the release of histamine induced by PEI, (3 pg/ml) from 41.1 + 2.0 (in the absence of LFA. n = 6) to 13.5 f 3.0 (in the presence of 100 pg/ml LFA, n = 6) of the total cell content. Moreover, the release of histamine induced by compound 48/80 was inhibited by WGA and PHA. but not by Con A, suggesting that the binding sites for PEI, and compound 48/80 had siak acid and GalNAc residues. The binding sites for PEI, seemed to especially overlap those of compound 48/80. Therefore, the binding of PEI, to their binding sites of mast cells seemed to activate G protein to release histamine from the mast cells. It is possible that mast cells also have other binding sites for PAAs, because WGA, PHA and Con A partially inhibited the histamine release induced by PAA with a molecular weight of 10,000. Thus PEIs and PAAs should be useful in further studies on the mechanisms of signal transduction coupling with G proteins of rat mast ce!!s.
711
References Suzu~-IWGmw T., SefiW H..Yoshind,Y., Nagaya, K.. Oku, N.. Nango, M. and Uchida, M.K., 1989, Jpn. N., Yamaguchi, N., Yamaguchi, N., Shibamoto, S., Ito. F. and Nango. M., 1986, J. B&hem. It-~o, 935.
J_ Ph~acol.51.279_
Oh
El
P.tu.012
relaxing factor an
a!st ee
i&amine release Mannaioni,
P.F., Masini, E., Salvemini
*, D., Pistelli, A., Korbut *, R. and Vane *, J.R.
Dept. oj Preciinical and Clinical Pharmacology, Viale G. B. Morgagni 65, 50134 Florence. Italy and * The WiU:am Harvey Research Institute, St. Bartholomew’s Hospital Medical College, Charterhouse Square. London EC JM 6 BQ, U.K.
Activated endothelial cells release a labile factor which relaxes vascular smooth muscle and inhibits platelet aggregation. This endothelium-derived relaxing factor (EDRF) is accounted for by nitric oxide (NO) synthetised from the amino-acid I-arginine, acting via stimulation of soluble guanylate cyclase and whose biological actions are terminated by superoxide anion and by oxy-haemoglobin (Moncada et al., 1989). Beside endothelial cells, relaxing factors have been shown to be released by activated neutrophils. macrophages. hepatocytes, Kupfer cells and neurons (Griffith et al., 1989). Here we show that isolated rat serosal mast cells ‘release a factor that inhibits p!atelet aggregation and has a pharmacological profile similar to that described for EDRF (NO). In turn, drugs which are capable of generaring NO inhibit mast cell histamine release induced by different stimuli. Incubation of mast cells with washed human platelets resulted in an inhibition of thrombine-induced platelet aggregation that was dependent on the number of the cells added. The inhibition was potentiated by superoxide dismutase and reversed by oxy-haemoglobin. Mast cells are more potent than bovine aortic endothelial cells in their ability to release a nitric oxide-like factor. A NO-generating drug (sodium nitroprusside. NaNP) was shown capable of inhibiting the release of histamine induced by compound 48/80, Ca2+ ionophore A 23187 and by a free radical-generating system (paracetamol+ prostaglandin-H-synthetase, PHS) in rat serosal mast cells. The inhibiting action was concentration-dependent. had a long time-course, was potentiated by the presence of an endotoxin lipopolysaccharide and was reversed by OXYhaemoglobin. These data show that the nitric oxide-like factor modulates the release of mast cell histamine. The work carried out at the William Harvey Res. Institute was supported by a grant from Glaxo Group Research Ltd. References Moncada, S., Palmer, R.M.J. and Higgs. E.A., 1989.
Biosynthesis
of nitric
oxide
from
L-Arginine.
1709-1715. Griffith,
T. and Randall,
M.. 1989. Nitric
oxide comes of age. The Lancet. October 7. 875-876.
Biochem.
Pharmacol.
38.