Journal of Cell Science 101, 745-750 (1992)
Printed in Great Britain © The Company of Biologists Limited 1992
745
Histamine decreases the permeability of an endothelial cell monolayer by
stimulating cyclic AMP production through the H2-receptor
TAICHI TAKEDA 12 , YUKO YAMASHITA1, SYUJI SHIMAZAKI2 and YOUJI MITSUI1*
1
Division of Cell Science and Technology, Fermentation Research Institute, Agency of Industrial Science and Technology, 1-1-3 Higashi,
Tsukuba city, lbaraki 305, Japan
2
Department of Traumatology and Critical Care Medicine, Kyorin University, 6-20-2 Shinkawa, Mitaka city, Tokyo 181, Japan
•Author for correspondence
Summary
To determine if histamine acts directly on the vascular
endothelium, the effect of histamine on the permeability
of cultured human endothelial cell monolayers and the
role of second messengers were examined. The addition
of 10~ 6 to 10~ 4 M histamine to the culture medium
decreased the endothelial cell monolayer permeability
and increased both cyclic AMP and free-calcium levels.
The decrease in permeability and the increase in cyclic
AMP mediated by histamine were prevented by an H2-
blocker (famotidine) while the increase in free-calcium
was inhibited by an Hj-blocker (diphenhydramine).
These results suggest that histamine decreases the
permeability of endothelial cell monolayers through the
H2-receptor, and cyclic AMP plays a more important
role than calcium ion as a second messenger.
Introduction
monolayer permeability to albumin while it increases
intracellular cyclic AMP through the H 2 -receptor.
Accompanying these changes was an increase in
cytoplasmic Ca 2 + via the Hi-receptor. However, Ca 2 +
seems not to play a major role in the permeability
change induced by histamine, since the Hi-blocker did
not inhibit the decrease of permeability caused by
histamine.
Vascular permeability is regulated by several humoral
factors such as histamine, bradykinin, catecholamines
and interleukins. In injured patients, histamine is
believed to increase vascular permeability (Matoltsy
and Matoltsy, 1951; Hayashi et al. 1964). In fact, many
investigators have described injury to endothelial cells
when histamine is administered in vivo (Matoltsy and
Matoltsy, 1951; Majno and Palade, 1961; Udaka et al.
1970; Gabbianti et al. 1970; Pietra et al. 1971;
Northover, 1975; McNamee and Grodins, 1975; Kaliner
et al. 1982). Histamine concentration is increased in the
interstitial fluid local to an injury and in the serum in the
early stages after injury (Horakova and Beaven, 1974;
Sampson and Archer, 1967). However, the direct
effects of histamine on the vascular wall are not
established. According to studies using cultured endothelial cells, histamine causes changes in cell shape
(Antonov et al. 1986), mediates cell mobility (Bottaro
et al. 1985), and accelerates cell growth (D'amore and
Shepro, 1977) and prostacyclin production (Baenziger
et al. 1980; Baenziger et al. 1981; Alhenc-Gelas et al.
1982). However, the permeability of the endothelial cell
layer has not been examined in the in vitro system. We
developed an in vitro model for assay of endothelial cell
monolayer permeability and examined the direct effect
of histamine on the endothelial cell monolayer. Our
results show that histamine decreases endothelial cell
Key words: endothelial cell monolayer, permeability,
histamine, cyclic AMP.
Materials and methods
Materials
MCDB151 was obtained from Sigma Chemical Co. (MO,
USA). Fetal bovine serum (FBS) was from Cell Culture
Laboratories Inc. (OH, USA), and was heat-inactivated at
56CC for 30 min. Endothelial cell growth substance (ECGS)
was purified from bovine brain and checked for its growthinducing activity (Imamura and Mitsui, 1987). Fibronectin
was a generous gift from Ito Ham (Japan). Most of the other
reagents were of special grades and manufactured by Wako
Pure Chemical Industries, Ltd. (Japan). The co-culture
chamber (Intercell) was purchased from Kurabou (Japan).
Preparation of cells and the monolayer system
Human umbilical vein endothelial cells (HUE147) were
isolated as described previously (Imamura and Mitsui, 1987),
and an endothelial cell line from a calf pulmonary artery was
designated as PACE2. Cells were maintained in MCDB151
supplemented with 15% FBS containing 2.5 ng/ml ECGS and
746
T. Takeda and others
5 /ig/ml heparin to maintain growth, and subcultured after
treatment with 0.25% trypsin. Cells were studied between
passages 10 and 20.
HUE147 and PACE2 cell lines in the growth phase were
seeded at a density of l ^ x ^ / c m 2 and ^xVf'/cm1, respectively, into co-culture chambers containing 200 fi\ of culture
medium. The Teflon membrane of the chamber was coated
with bovine fibronectin at 10 jig/cm2. One chamber was placed
in each well of a 24-well plate containing 600 /A of the same
culture medium. To prepare a confluent cell monolayer, cells
were allowed to proliferate on the Teflon membrane for 2
days at 37°C in a humidified incubator under a 95% :5% (air:
CO2) atmosphere.
10~6 10"
10
histamine (M)
10"
Permeability studies
Histamine, diphenhydramine hydrochloride and famotidine
were dissolved in MCDB151 + 15% FBS at various
concentrations. A 600 /d sample of each assay medium
containing 2 ^M FITC-conjugated bovine albumin was added
gently to each chamber and then incubated. After 2 hours the
medium in the lower chamber was removed and the
fluorescence intensity was measured with a fluorescence
spectrophotometer (Shimazu, Japan) (excitation, 490 nm;
emission, 525 nm).
Assay of cyclic AMP
Human umbilical vein endothelial cells (HUE147) were
seeded intofibronectin-coated6-well plates and cultured for 2
days. The cells were then exposed to 1 ml of fresh assay
medium containing 10~5 M histamine and/or 10~6 M
diphenhydramine or 10~8 M famotidine. After incubation for
10 min, the medium was removed to assay for cyclic AMP.
Cyclic AMP was measured in fmol/cm2 per well using a cyclic
AMP [125I]RIA kit (Amersham, UK) (non-acetylation protocol).
Determination of cytoplasmic Ca2+
HUE147 endothelial cells were incubated with culture
medium containing 2 /JM Indo-1 AM (a calcium-sensitive
fluorescent probe) for 20 min. The cells were then washed
twice gently with MCDB151 and exposed to assay medium
containing histamine and/or diphenhydramine or famotidine.
Ca2+ -bound and free Indo-1 in each cell were excited with a
351-363 nm argon ion laser and the immunofluorescence was
measured at 405 nm and 485 nm, respectively, using an
ACAS470 fluorimeter (Meridian Instruments, MI, USA).
Statistical analysis
All values are expressed as the mean ± s.d. Comparisons of
data in the permeability study and cyclic AMP assays were
made by one-way ANOVA and Bonferoni's method. Comparisons of data from the Ca2+ assay were assessed by the chisquared test after Yates' correction. Variations were significant at the P<0.05 level.
Results
Both human umbilical vein and calf pulmonary artery
endothelial cells formed a confluent monolayer on the
10 /zg/cm2 fibronectin-coated Teflon membrane of a coculture chamber (as verified by phase-contrast and
scanning electron microscopy). In the confluent human
umbilical vein endothelial cell monolayer the permeability to albumin was decreased to 15-25% of the
control containing membrane only. The permeability of
0 10"8 10
10"5 10~4 10"
histamine (M)
Fig. 1. Permeability of endothelial cell monolayers to
FITC-albumin at different concentrations of histamine.
Histamine was applied to the upper chamber of (A) human
umbilical vein endothelial cell (HUE147) monolayer and
(B) bovine pulmonary artery endothelial cell (PACE2)
monolayer in various concentrations and the permeability
of the monolayer to FITC-albumin was examined. The
difference in the decrease in permeability was significant
between3 10~6 and 10~4 M histamine in HUE147, and 10~5
and 1(T M histamine in PACE2 (F<0.05). Data are
expressed as the mean ± s.d. (n=4).
a confluent monolayer of pulmonary artery endothelial
cells was decreased to 60-80% of the control. Fibronectin coating alone did not affect permeability. The
permeability to albumin was measured after 2 h when
FITC-albumin reaches a detectable concentration
under our assay system.
Histamine at a concentration of 10~6 to 10~4 M
decreased the permeability of the membrane to albumin in a culture of HUE147 cells (Fig. 1 A). The
permeability of PACE2 cells to albumin was also
decreased by histamine in a dose-dependent manner
(Fig. 1 B). In both cases, the permeability decreased
but reached a lower plateau level, and in HUE147 cells
it even began to increase, at higher doses of histamine.
The decrease in permeability of HUE147 cells to
albumin in the presence of 10~5 M histamine was
blocked by famotidine (Fig. 2 A). Diphenhydramine,
however, did not have any statistically significant effect
even at the high concentration of 10~4 M (Fig. 2 B).
Histamine also increased cyclic AMP production in a
dose-dependent manner and reached a maximum at 10
min after its addition to cells (data not shown). Fig. 3
shows that 10~8 M famotidine reduced histaminestimulated cyclic AMP production but treatment with
Histamine and endothelial cell permeability
747
A
HI
"8
icr5 (M)
iiiaiaiuiiii'
10"6 (M)
famotidine
10~5 (M)
10"4 (M)
nisiaminc
diphenhydramine
500 ms
Fig. 2. Permeability of endothelial cell monolayers to
FITC-albumin in the presence of 10~5 M histamine and
different concentrations of famotidine or diphenhydramine.
(A) 10~5 M histamine and various concentrations of
famotidine (H2-blocker) were applied and the FITCalbumin permeability of the endothelial cell (HUE147)
monolayer was examined. 10~8 to 10~6 M famotidine
inhibited the decrease in permeability caused by histamine
(P<0.05). (B) 10~5 M histamine and various
concentrations of diphenhydramine (Hrblocker) were
applied. 10~6 to 10~4 M diphenhydramine did not affect
the permeability (P<0.05). Data are expressed as the mean
± s.d. (n=4).
Fig. 4. Ca2+ concentration in endothelial cells as
determined by ACAS. 10~4 M diphenhydramine and 10~8
M famotidine were applied in the presence of 10~5 M
histamine (arrow) to endothelial cells (HUE147). Typical
examples are shown. (A) Histamine increased the Ca2+
concentration in endothelial cells in two phases. In the
initial phase, histamine increased Ca2+ rapidly to 50-150
nM. Later the increase in Ca2+ concentration seen with
histamine oscillated continuously over a 40 nM range.
(B) 10~4 M diphenhydramine applied simultaneously with
histamine inhibited later oscillations in the Ca2+
concentration. (C) 10~8 M famotidine did not affect the
histamine-induced Ca2+ increase.
control
1CT4 M diphenhydramine did not affect the cyclic AMP
level.
The cytoplasmic Ca 2+ concentration of isolated
endothelial cells increased rapidly with 10~5 M histamine (Fig. 4 A), regardless of the presence of 10~8 M
famotidine (Fig. 4 C: for experimental details see
legend). In contrast, 10~4 M diphenhydramine, the
concentration used in the permeability studies and
assay of cyclic AMP, inhibited the oscillations in Ca 2+
concentration after the initial rapid increase (Fig. 4 B).
Neither famotidine nor diphenhydramine themselves
had any effect on the cytoplasmic Ca 2+ concentration.
Pretreatment with diphenhydramine 20 min prior to
adding histamine inhibited both the initial rapid
increase and the latter fluctuations in cytoplasmic Ca 2+
concentration. Inhibition of the initial rise was observed
only after preincubation, although diphenhydramine is
a receptor blocker. It has not been determined why
diphenhydramine was not immediately effective when
applied to cells in culture. Famotidine pretreatment did
not alter this effect. This is illustrated in Table 1, which
shows the effects of histamine and each antagonist on
1 /><0.05
10~5 M histamine
10~4 M diphenhydramine
10~4 M diphenhydramine
+ 10~5 M histamine
8
10~ M famotidine
1(T8 M famotidine
+ 10~5 M histamine
P<0.05
0
20 40
cyclic AMP (fmol/cm2)
60
Fig. 3. Cyclic AMP content of endothelial cells determined
by radioimmunoassay. 10~5 M histamine and/or 10~4 M
diphenhydramine or 10~5 M histamine and/or 10~8 M
famotidine were applied for 10 min to an endothelial cell
(HUE147) monolayer. Phosphate buffered saline was
applied as a control. Histamine increased cyclic AMP in
endothelial cells. 10~4 M diphenhydramine did not inhibit
the increase in cyclic AMP caused by 10~5 M histamine.
10~8 M famotidine inhibited this histamine-induced
increase in cyclic AMP. Data are epressed as the mean ±
s.d. (n=3). Differences noted were significant at P<0.05.
748
T. Takeda and others
Table 1. The frequency of the initial rapid rise and subsequent oscillations in Ca2+ concentration in endothelial
cells (HUE147) determined for each type of experiment
Number and percentage
of positive occurrences
Applied drugs
Number of
experiments
Initial rise (%)
12
25
10
40
10
0(0)
21(84)
0(0)
31 (76)
9(90)
10
13
10
0(0)
12(92)
0(0)
Control
10~5 M histamine
10"8 M famotidine
10~5 M histamine + 10~8 M famotidine
10"5 M histamine + 10"8 M famotidine
(pretreatment)
10~4 M diphenhydramine
5
10~ M histamine + 10~4 M diphenhydramine
10~! M histamine + 10~4 M diphenhydramine
(pretreatment)
Subsequent
oscillations (%)
0(0)
20(80)
0(0)
27 (63)*
4 (44)
0(0)
0(0)t
0(0)
The number and percentage of positive occurrences in each jjhase are shown. 10~5 M'. histaminecaused both an initial rapid rise and
subsequent oscillations. 10 M famotidine did not affect these increases in Ca2+ concentration. 10 ~4 M diphenhydramine applied
simultaneously with histamine inhibited the subsequent Ca2+ concentration oscillations. Pretreatment with 10 M diphenhydramine for 20
min inhibited both the initial rise and the subsequent oscillations in the cytoplasmic Ca2+ concentration.
*Not different from 10~5 M histamine.
tDifferent from 10"5 M histamine (P<0.05).
the frequency of the initial rapid rise and subsequent
oscillations of cytoplasmic Ca that were observed in a
large number of experiments. Pretreatment with famotidine, in contrast to diphenhydramine, did not affect
the histamine-induced large rise in cytoplasmic Ca2+
but inhibited, somewhat, subsequent Ca oscillations.
Discussion
The effects of histamine on endothelial cells are thought
to be mediated by the histamine receptors on these cells
(Ash and Schild, 1966; Buonassi and Venter, 1976;
Berti et al. 1979; Baenziger et al. 1980; Simionescu et al.
1982; Van de Voorde and Leusen, 1983). The receptors
can be subdivided into two classes, Hi and H2, based on
their physiology and pharmacology, and on the second
messengers specific to each. It is reported that the
second messengers of histamine via the Hi-receptor are
phosphatidylinositol, diacylglycerol and calcium, and
via the H2-receptor it is cyclic AMP (Grigorian et al.
1989). In this study, we show that the histamine-induced
decrease in the permeability of an endothelial cell
monolayer to albumin is accompanied by an increase in
the cyclic AMP concentration of endothelial cells.
These effects of histamine were inhibited by an H2blocker (famotidine) and not by an Hi-blocker (diphenhydramine). Histamine also increases the concentration
of Ca2+ in endothelial cells. This effect, however, was
inhibited by the Hj-blocker but not by the H2-blocker.
On the basis of these observations we suggest that
histamine alters the permeability of the endothelial cell
monolayer via H2-receptors and cyclic AMP and that
the cytoplasmic Ca2+ concentration does not affect cell
permeability.
Histamine is reported to stimulate endothelial cell
proliferation (D'Amore and Shepro, 1977). Because
the population doubling time of HUE 147 is 22 hours
and our results were obtained over a two-hour period,
the histamine-induced decrease in permeability cannot
be due to an increase in the number of cells. MizunoYagyu et al. (1987) reported that PGI2 (prostocyclin)
inhibited dextran transport through the endothelial cell
monolayer and that this effect was mediated by
increased cyclic AMP production. On the other hand,
Baenziger et al. (1980) showed that histamine stimulates the production of PGI2 via an Hi-receptormediated mechanism. Our data, however, support the
belief that histamine itself decreases the permeability of
the endothelial cell monolayer primarily through the
H2-receptor. This suggests that the decrease in permeability due to histamine does not require PGI2
synthesis.
Regarding the role of cyclic AMP, it has been
reported that membrane-permeable analogues of cyclic
AMP, 8-bromo cyclic AMP and dibutyryl cyclic AMP
decrease the permeability of the endothelial cell
monolayer (Casnocha et al. 1989). Duffey et al. (1981)
reported that a correlation exists between electrical
resistance and the cyclic AMP content of epithelial
cells. Since albumin passes through the endothelial cell
layer intercellularly (junctional way) and/or transcellularly (vesicular transport) (Navab et al. 1986), cyclic
AMP may increase the number of junctions between
endothelial cells and narrow the intercellular space.
Synchronized cytoplasmic Ca 2+ oscillations have
been reported in confluent monolayers of human
endothelial cells (Sage et al. 1989; Neylon and Irvine,
1990). Neylon and Irvine reported that the synchronized repetitive spikes in cytoplasmic calcium occur in
response to histamine in confluent human umbilical
vein endothelial cell monolayers, and that spiking
behavior is not seen in non-confluent cell monolayers.
In this study, we observed the oscillations in Ca 2+
concentration in single cells using an ACAS470 fluorimeter. The cells were spread on a coverglass sparsely
Histamine and endothelial cell permeability
and had no contact between each other. However, the
repetitive spikes in a single cell are still evoked by
histamine, and these spikes are inhibited by diphenhydramine (Hj-blocker). At present it is not clear
whether these oscillations are generated by the same
mechanism as those seen in monolayer cells.
Several studies have shown that histamine increases
vascular permeability in vivo (Matoltsy and Matoltsy,
1951; Hayashi et al. 1964). We now report that
histamine decreases endothelial cell monolayer permeability in vitro. Our in vitro system of an endothelial
monolayer was designed to represent a simple model of
capillary vessels. There always remains the question of
its relevance to the situation in vivo. Albelda et al.
(1988) reported that the calculated permeability of
albumin across an in vitro monolayer is 10-100 times
higher than that found in vivo, although they used fetal
bovine aortic endothelial cells. The reasons for the
differences are not known. The authors observed
occasional gaps between adjacent cells (5-10%) in their
in vitro model, and also found a lack of charge
selectivity. The influence of the basement membrane
and/or interactions between endothelial cells and other
cells or humoral factors in vivo would also result in
differences between the intact endothelium and in vitro
models. However, we believe that the use of an in vitro
model for permeability has a number of advantages,
such as offering direct access to luminal and abluminal
fluid for analysis and being highly simplified and limited
to a single cell type. Our results suggest that the direct
effect of histamine on the endothelial cell monolayer is
to decrease permeability and that when histamine
increases the permeability, it does so indirectly by
stimulating the surroundings in vivo.
The authors thank Dr. A. Iwashima for advice on the Ca2+
assay.
This work was supported by a project grant for Basic
Technology for Future Industry from the Ministry of
International Trade and Industry of Japan.
Data in Fig. 1 A were presented at the 18th Annual Meeting
of the Japanese Association for Acute Medicine, Kurashiki
city, Japan, November 9, 1990.
References
Albelda, S. M., Sampson, P. M., Haselton, F. R., McNifT, J. M.,
Mueller, S. N., Williams, S. K., Flshman, A. P. and Levlne, E. M.
(1988). Permeability characteristics of cultured endothelial cell
monolayers. J. Appl. Physiol. 64, 308-322.
Alhenc-Gelas, F., Tsai, S. J., Callahan, K. S., Campbell, W. B. and
Johnson, A. R. (1982). Stimulation of prostaglandin formation by
vasoactive mediators in cultured human endothelial cells.
Prostaglandins 24, 723-742.
Antonov, A. S., Lukashev, M. E., Romanov, Y. A., Tkachuk, V. A.,
Repln, V. S. and Smirnov, V. N. (1986). Morphological alterations
in endothelial cells from human aorta and umbilical vein induced by
forskolin and phorbol 12-myristate 13-acetate: A synergistic action
of adenylate cyclase and protein kinase C activators. Proc. Nat.
Acad. Sci. U.S.A. 83, 9704-9708.
Ash, A. S. F. and Schlld, H. O. (1966). Receptors mediating some
actions of histamine. Brit. J. Pharmacol. Chemother. 27, 427-439.
BaenzJger, N. L., Fogerty, F. J., Mertz, L. F. and Chernuta, L. F.
(1981). Regulation of histamine-mediated prostacyclin synthesis in
cultured human vascular endothelial cells. Cell 24, 915-923.
749
Baenziger, N. L., Force, L. E. and Becherer, P. R. (1980). Histamine
stimulates prostacyclin synthesis in cultured human umbilical vein
endothelial cells. Biochem. Biophys. Res. Commun. 92, 14351440.
Berti, F., Folco, G. C , Nicosia, S., Omlnl, C. and Pasarglkllan, R.
(1979). The role of histamine Hi- and H 2 -receptors in the
generation of thromboxane A 2 in perfused guinea-pig lungs. Brit.
J. Pharmacol. 65, 629-633.
Bottaro, D., Shepro, D., Peterson, S. and Hechtman, H. B. (1985).
Serotonin, histamine, and norepinephrine mediation of endothelial
and vascular smooth muscle cell movement. Amer. J. Physiol. 248,
C252-C257.
Buonassisi, V. and Venter, J. C. (1976). Hormone and
neurotransmitter receptors in an established vascular endothelial
cell line. Proc. Nat. Acad. Sci. U.S.A. 73, 1612-1616.
Casnocha, S. A., Eskln, S. G., Hall, E. R. and Mclntire, L. V. (1989).
Permeability of human endothelial monolayers: effect of vasoactive
agonists and cAMP. /. Appl. Physiol. 67, 1997-2005.
D'Amore, P. and Shepro, D. (1977). Stimulation of growth and
calcium influx in cultured, bovine, aortic endothelial cells by
platelets and vasoactive substances. /. Cell. Physiol. 92, 177-184.
DufTey, M. E., Hainau, B., Ho, S. and Bentzel, C. J. (1981).
Regulation of epithelial tight junction permeability by cyclic AMP.
Nature 294, 451-453.
Gabblanti, G., Badonnel, M. C. and Majno, G. (1970). Intra-arterial
injections of histamine, serotonin, or bradykinin: A topographic
study of vascular leakage. Proc. Soc. Exp. Biol. Med. 135, 447-452.
Grigorian, G. Y., Mirzapoyazova, T. Y., Resink, T. J., Danilov, S. M.
and Tkachuk, V. A. (1989). Regulation of phosphoinositide
turnover in endothelium from human pulmonary artery, aorta and
umbilical vein. Antagonistic action on the /3-adrenoceptor coupled
adenylate cyclase system. J. Mol. Cell. Cardiol. 21, 119-123.
Hayashi, H., Yoshlnaga, M., Koono, M., Mlyoshl, H. and
Matsumura. M. (1964). Endogeneous permeability factors and
their inhibitors affecting vascular permeability in cutaneous arthus
reactions and thermal injury. Brit. J. Exp. Pathol. 45, 419-435.
Horakova, Z. and Beaven, M. A. (1974). Time course of histamine
release and edema formation in the rat paw after thermal injury.
Eur. J. Pharmacol. 27, 305-312.
Imamura, T. and Mltsni, Y. (1987). Heparan sulfate and heparin as a
potentiator or a suppressor of growth of normal and transformed
vascular endothelial cells. Exp. Cell Res. 172, 92-100.
Kaliner, M., Shelhamer, J. H. and Ottesen, E. A. (1982). Effects of
infused histamine: correlation of plasma histamine levels and
symptoms. /. Allergy Clin. Immunol. 69, 283-289.
Majno, G. and Palade, G. E. (1961). The effect of histamine and
serotonin on vascular permeability: An electron microscopic study.
/. Biophys. Biochem. Cytol. 11, 571-606.
Matoltsy, A. G. and Matoltsy, M. (1951). The action of histamine and
antihistaminic substances on the endothelial cells of the small
capillaries in the skin. J. Pharmacol. Exp. Ther. 102, 237-249.
McNamee, J. E. and Grodins, F. S. (1975). Effect of histamine on
micTovasculature of isolated dog gracilis muscle. Amer. J. Physiol.
229, 119-125.
Mizuno-Yagyn, Y., Hashida, R., Mlneo, C , Ikegami, S., Ohkuma, S.
and Takano, T. (1987). Effect of PGI 2 on transcellular transport of
fluorescein dextran through an arterial endothelial monolayer.
Biochem. Pharmacol. 36, 3809-3813.
Navab, M., Hough, G. P., Berliner, J. A., Frank, J. A., Fogehnan, A.
M., Haberland, M. E. and Edwards, P. A. (1986). Rabbit betamigrating very low density lipoprotein increases endothelial
macromolecular transport without altering electrical resistance. /.
Clin. Invest. 78, 389-397.
Neylon, C. B. and Irvine, R. F. (1990). Synchronized repetitive spikes
in cytoplasmic calcium in confluent monolayers of human umbilical
vein endothelial cells. FEBS Lett. 275, 173-176.
Northover, A. M. (1975). Action of histamine on endothelial cells of
guinea-pig isolated hepatic portal vein and its modification by
indomethacin or removal of calcium. Brit. J. Exp. Pathol. 56, 5261.
Pietra, G. G., Szidon, J. P., Leventhal, M. M. and Flshman, A. P.
(1971). Histamine and interstitial pulmonary edema in the dog.
Circul. Res. 29, 323-337.
Sage, S. O., Adams, D. J. and van Breemen, C. (1989). Synchronized
750
T. Takeda and others
oscillations in cytoplasmic free calcium concentration in confluent
bradykinin-stimulated bovine pulmonary artery endothelial cell
monolayers. J. Biol. Chem. 264, 6-9.
Sampson, D. and Archer, G. T. (1967). Release of histamine from
human basophils. Blood 29, 722-736.
Simionescu, N., Heltianu, C , Antohe, F. and Slmionescu, M. (1982).
Endothelial cell receptors for histamine. Ann. N. Y. Acad. Sci. 401,
132-149.
Udaka, K., Takenchl, Y. and Movat, H. Z. (1970). Simple method for
quantitation of enhanced vascular permeability. Proc. Soc. Exp.
Biol. Med. 133, 1384-1387.
Van de Voorde, J. and Leusen, I. (1983). Role of the endothelium in
the vasodilator response of rat thoracic aorta to histamine. Eur. J.
Pharmacol. 87, 113-120.
(Received 5 August 1991 - Accepted, in revised form,
14 January 1992)