(Circulation. 1996;93:161-167.)
© 1996 American Heart Association, Inc.
Articles |
Diabetes Exacerbates Inflammatory Responses to Ischemia-Reperfusion
From the Department of Physiology, Louisiana State University Medical Center, Shreveport; the Discovery Research, UpJohn Laboratories, Kalamazoo, Mich (D.C.A.); and the Department of Bioregulation, Biomedical Research Center, Osaka University Medical School, Japan (M.M.).
Correspondence to D. Neil Granger, PhD, Department of Physiology, LSU Medical Center, 1501 Kings Hwy, PO Box 33932, Shreveport, LA 71130-3932. E-mail dgrang@lsumc.edu.
 |
Abstract |
|---|
 Top Abstract IntroductionMethods Results Discussion References |
|---|
Background Diabetes is associated with an increased incidence of ischemic organ damage. The objectives of present study were to compare the leukocyte–endothelial cell adhesive interactions and albumin leakage response of mesenteric venules to ischemia-reperfusion between control rats, rats with streptozotocin-induced diabetes, and rats with hyperglycemia induced by glucose infusion and to define the molecular determinants of the leukocyte accumulation elicited by ischemia-reperfusion in diabetic rats.
Methods and Results Under baseline conditions, lower venular shear rates and an increased number of rolling leukocytes were noted in diabetic rats, whereas the number of adherent and emigrated leukocytes did not differ from that in control rats. Spontaneous albumin leakage from mesenteric venules was markedly increased in diabetic rats but not in hyperglycemic nondiabetic rats. Ischemia-reperfusion elicited significantly larger increases in leukocyte adhesion and emigration and albumin leakage in diabetic rats. Acute elevation of glucose levels did not modify the microvascular responses to ischemia-reperfusion compared with control rats. Antibodies directed against CD11/CD18, intercellular adhesion molecule–1 (ICAM-1), or P-selectin but not L-selectin significantly decreased the number of adherent and emigrated leukocytes after ischemia-reperfusion in diabetic rats. However, none of the antibodies significantly attenuated the increased albumin leakage response to ischemia-reperfusion in diabetic rats.
Conclusions Thes results indicate that diabetes mellitus is associated with exaggerated leukocyte–endothelial cell adhesion and albumin leakage responses to ischemia-reperfusion. The enhanced leukocyte accumulation in response to ischemia-reperfusion is mediated by CD11/CD18–ICAM-1 interactions (firm adhesion) and P-selectin (rolling). The exaggerated albumin leakage response to ischemia-reperfusion in diabetics is not mediated by the recruited inflammatory cells.
Key Words: ischemia • reperfusion • leukocytes • microcirculation • risk factors
|
Introduction |
|---|
|
TopAbstractIntroductionMethods Results Discussion References |
|---|
Accelerated ischemic vascular disease is the principal cause of mortality in patients with diabetes mellitus.1 Hyperglycemia, hypertension, and abnormalities in lipid and lipoprotein metabolism are risk factors that may predispose the diabetic vasculature to the deleterious consequences of ischemia and reperfusion. However, none of these cardiovascular risk factors, either alone or in combination, appear to fully account for the marked increase in mortality associated with diabetes.2
Over the past decade, a large body of evidence has accumulated that implicates neutrophils in the pathobiology of different cardiovascular diseases, including hypertension,3 atherosclerosis,4 stroke,5 and myocardial ischemia.6 A role for neutrophils in these cardiovascular diseases has been invoked on the basis of studies demonstrating that (1) neutrophils are activated and accumulate in affected tissues, (2) neutrophil depletion attenuates the accompanying microvascular and parenchymal cell dysfunction, and (3) the extent of tissue damage and organ dysfunction is reduced by agents that prevent the activation or accumulation of neutrophils.7 8 9 10 One means of preventing leukocyte sequestration that has proved to be very effective is immunoneutralization of adhesion glycoproteins expressed on the surface of leukocytes or endothelial cells.11 Monoclonal antibodies (mAbs) directed against these molecular determinants of leukocyte–endothelial cell adhesion have been shown to afford protection in a variety of experimental models of ischemic disease.8 9 10
A growing body of evidence also shows that the function and mechanical properties of granulocytes are altered in diabetes. Granulocytes isolated from diabetic animals and humans are less deformable12 13 14 and generate larger quantities of toxic oxygen radicals15 than granulocytes isolated from nondiabetics. Moreover, a larger percentage of the circulating neutrophils is activated in human diabetics compared with control populations.16 Taken together, these observations suggest that the activated and less deformable polymorphonuclear leukocytes may predispose diabetics to neutrophil-mediated tissue injury. Although it would appear likely that diabetes predisposes tissues to the deleterious consequences of ischemia and reperfusion, an injury process that is largely mediated by neutrophils in some tissues, there is no experimental evidence that directly addresses this important issue. Thus, the overall objective of this study was to determine whether diabetes renders the microvasculature more vulnerable to the deleterious effects of ischemia-reperfusion. In addition, we addressed the possibility that an enhanced accumulation of adherent leukocytes within postcapillary venules or hyperglycemia per se contributes to the microvascular dysfunction observed in diabetic tissues exposed to ischemia-reperfusion.
|
Methods |
|---|
|
TopAbstractIntroductionMethods Results Discussion References |
|---|
Animal Model
Male Sprague-Dawley rats weighing 200 to 225 g were obtained from Harlan. After a 5-day adaptation period, diabetes mellitus was induced by injection of streptozotocin (65 mg/kg IP; Sigma Chemical Co)17 diluted in citrate buffer at a concentration of 6.5 mg/mL. Control rats received an intraperitoneal injection of the corresponding amount of citrate buffer alone. Induction of the diabetic condition, defined as a serum glucose concentration >200 mg/dL, was confirmed by glucose measurements in serum samples obtained from the tail vein at 1 and 3 weeks (520±27 and 480±35 mg/dL, respectively) after streptozotocin administration. All studies were performed 3 weeks after treatment, preceded by a 24-hour fasting period with free access to water. The experimental protocols were approved and performed in accordance with the guidelines of the Louisiana State University Medical Center Animal Care and Use Committee, Shreveport.
To assess the effects of acutely increased glucose levels on the microvascular responses to ischemia-reperfusion, two groups of previously untreated control rats were studied. In one group, the region of the mesentery under study was superfused with bicarbonate buffer containing glucose at a concentration of 480 mL/dL, which corresponds to the mean value of plasma glucose in the diabetic group at the time of study. In a second series of experiments, acute hyperglycemia was induced by continuous infusion of 50% glucose at 200 mg·kg-1·min-1 IV during the first 5 minutes and 75 mg·kg-1·min-1 thereafter.
Surgical Techniques
The animals were anesthetized with
thiobutabarbital
(Inactin) 90 mg/kg IP in nondiabetic rats and 60 mg/kg IP in diabetic
rats. A tracheotomy was performed on each rat to facilitate breathing
throughout the experiment, and the right carotid artery and right
jugular vein were cannulated. Systemic arterial pressure
was measured with a Statham P23A pressure transducer and recorded
with a polygraph DC driver amplifier (Grass Instrument Co). A midline
abdominal incision was made to allow for exteriorization of a section
of the mesentery from the small intestine. A blood sample was obtained
before the beginning of the study to determine total white blood cell
and neutrophil counts and plasma glucose concentration.
Intravital Microscopy
The rats were positioned on a
20x30-cm Plexiglas board in a
manner that allowed a selected section of mesentery to be placed over a
glass slide covering a 3.5x3.5-cm hole centered in the Plexiglas. All
exposed tissue was covered with saline-soaked gauze to minimize
tissue dehydration. The board was mounted onto the stage of an
orthostatic microscope (Nikon Optihot). The image produced
with a x40 objective (Nikon E-Plan 40) was captured on videotape
(BR–S601MU videocassette recorder, JVC) with a color camera
(VK-C150, Hitachi). The time and date were displayed on both taped and
live images (Trinitron monitor, Sony) with a date-time generator
(WJ-810, Panasonic). The mesentery was superfused at 2.5 mL/min with
bicarbonate-buffered saline bubbled with a 95%
N2/5% CO2 gas mixture to reduce the
oxygen tension to the physiological
intraperitoneal level (40 to 50 mm Hg). The
superfusate was maintained at 37°C by pumping the
solution through a heat exchanger warmed with a
constant-temperature circulator (model 801, Fisher Scientific).
Rectal and mesenteric temperatures were monitored with an
electrothermometer. Body temperature was kept between 36.5°C and
37.5°C with an infrared heat lamp.
Single unbranched venules with diameters of 25 to 35 µm and lengths >150 µm were selected for study. Venular diameter (DV) was measured on-line with a video caliper (Microcirculation Research Institute, Texas A&M University, College Station). The number of adherent, emigrated, and rolling leukocytes was determined off-line during playback of videotaped images. A leukocyte was considered adherent to venular endothelium if it remained stationary for 30 seconds or longer. Adherent leukocytes were quantified as the number per 100-µm length of venule. Leukocyte emigration was expressed as the number per microscopic field (2.12x10-2 mm2). Rolling leukocytes were defined as those white blood cells that moved at a velocity less than that of erythrocytes in the same vessel. Leukocyte rolling velocity (VLR) was determined from the time required for a leukocyte to traverse a 50-µm distance along the length of the venule and is expressed as micrometers per second. The flux of rolling leukocytes was measured as those white cells that could be seen moving within a small (10-µm) viewing area of the vessel with the same area used throughout the experiment. The number of rolling leukocytes per 100-µm venule length was calculated as the leukocyte flux divided by VLR.
Centerline red blood
cell velocity (VRBC) was measured with
an optical Doppler velocimeter (Microcirculation
Research Institute, Texas A&M University) that was calibrated against a
rotating glass disk coated with red cells. Venular blood flow was
calculated from the product of mean red blood cell velocity
(Vmean=Centerline Velocity/1.6) and microvascular
cross-sectional area, with cylindrical geometry assumed. Venular
wall shear rate (
) was calculated from the newtonian definition:
=8(Vmean/Dv).
To quantify albumin leakage across mesenteric venules, 25 mg/kg IV FITC-labeled rat albumin (Sigma Chemical Co) was administered to the rats. Fluorescence intensity (excitation wavelength, 420 to 490 nm; emission wavelength, 520 nm) was detected with a CCD camera model XC-77 (Hamamatsu Photonics), a C2400-60 CCD camera control unit, and a C2400-68 intensifier head (Hamamatsu Photonics) attached to the camera. The fluorescence intensity of the venule under study (Iv), the fluorescence intensity of contiguous perivenular interstitium within 10 to 50 µm of the venular wall (Ii), and the background fluorescence before injection of FITC-albumin (Ib) were analyzed with a Macintosh Quadra computer (Apple Computer Inc) equipped with a 24STV graphics display board (Rasterops Co) with the public-domain NIH IMAGE computer-assisted digital imaging processor. An index of vascular albumin leakage (permeability index) was determined according to the formula (Ii-Ib)/(Iv-Ib). Measurements of Iv and Ii were obtained at least 30 minutes after FITC-albumin injection.
Experimental Protocols
After a stabilization period of 30
minutes, images from the
mesenteric preparation were recorded on videotape for 10 minutes.
Thereafter, ischemia was induced by placing a ligature of
polyethylene P-50 tubing around the superior mesenteric artery and a
piece of polyethylene P-280 tubing around the superior mesenteric vein.
Ischemia was maintained for 10 minutes; then the P-280 tubing
was removed, allowing blood to recirculate. Only rats in which ligation
of the superior mesenteric vessels induced a decrease in centerline red
blood cell velocity 
80% were used in the study. All
parameters were measured again 5 to 15 minutes and 30 to 40
minutes after reperfusion. In some experiments, animals received mAbs
directed against the ß subunit (CD18) of CD11/CD18 (mAb CL26, 100
µg per rat),18 intercellular adhesion molecule–1
(ICAM-1; mAb 1A29, 2.0 mg/kg),19 L-selectin
(HLR3, 1 mg/kg),20 P-selectin (PB1.3, 2
mg/kg),21 or a nonbinding antibody (PNB1.6, 2.0
mg/kg),22 with the mAb administered immediately after the
baseline measurements were obtained (10 minutes before ischemia
was induced). The concentration of mAbs used in this study was based on
experiments that determined the amount of mAb needed to maximally
reduce leukocyte adherence and emigration in rat mesenteric venules
induced by inflammatory mediators23 or
ischemia-reperfusion.10
In the experiments designed to assess the effects of acutely increased glucose levels on the microvascular response to ischemia-reperfusion, glucose was either superfused directly onto the mesentery or infused intravenously (as described above) after the baseline measurements were obtained. Thirty minutes after the onset of exposure to high glucose levels, all parameters were measured again. Thereafter, ischemia-reperfusion was induced with the same protocol as described previously.
Statistical Analysis
The data were analyzed with standard
statistical
analyses, ie, ANOVA with Scheffé's (post hoc) test and
Student's paired or unpaired t test where appropriate. All
values are reported as mean±SEM from six to eight rats. Statistical
significance was set at P<.05.
|
Results |
|---|
|
TopAbstractIntroductionMethods Results Discussion References |
|---|
Under baseline conditions, diabetic rats had significantly lower blood pressure than control rats (104±4 versus 119±2 mm Hg; P<.05). Diabetic and control rats had very similar total white cell (7.1±0.7 versus 7.9±0.6 cells/mm3) and neutrophil (2.4±0.5 versus 2.5±0.5 cells/mm3) counts in blood. A proliferation of small blood vessels (10 to 50 µm in diameter) was observed in the mesentery of diabetic rats. The diameters of the venules studied were similar in diabetic and control rats (34±1 versus 33±1 µm). Mesenteric venules of diabetic rats had a significantly lower centerline red blood cell velocity (2.50±0.15 versus 3.68±0.39 mm/s, P<.01) and shear rate (368±29 versus 555±52 s-1, P<.01) than control rats. In diabetic rats, the basal leukocyte rolling velocity was significantly lower and the number of rolling leukocytes was significantly higher compared with control rats (Fig 1
). The decrease in rolling velocity was determined
largely by a reduction in blood flow velocity because after
normalization of leukocyte rolling velocity to the corresponding red
blood cell velocity (VLR/VRBC), no
significant differences were observed between control and diabetic rats
(0.016±0.002 versus 0.013±0.001). Under baseline conditions,
control
and diabetic rats had similar numbers of adherent and emigrated
leukocytes (Fig 2), whereas diabetic rats exhibited a
markedly increased albumin leakage relative to control rats
(39.0±8.7 versus 9.7±2.3%, P<.01; Fig
3).
|
|
|
During the ischemic period, centerline red blood cell velocity (0.43±0.06 versus 0.46±0.06 mm/s, P>.05) and shear rate (60.3±9.0 versus 69.5±11.1 s-1, P>.05) were similar in both groups of rats. After reperfusion, diabetic rats again exhibited a significantly lower centerline red blood cell velocity (1.52±0.22 versus 2.7±0.24 mm/s, P<.01) and shear rate (221±36 versus 406±27 s-1) than control rats. In both groups of rats, the values of red blood cell velocity and shear rate were significantly lower after reperfusion compared with values obtained under baseline conditions (P<.05, paired t test).
Diabetic rats exhibited a significant reduction in leukocyte rolling
velocity and significant increases in the flux of rolling leukocytes
and the number of rolling leukocytes immediately after reperfusion (Fig
4). Control (nondiabetic) rats exhibited similar
increases in the flux and number of rolling leukocytes, but leukocyte
rolling velocity was not significantly altered (Fig 4). The
alterations
in leukocyte rolling induced by ischemia-reperfusion were
short-lived and had returned entirely to control levels 30 minutes
after reperfusion (Fig 4). Ischemia-reperfusion caused a
significant increase in the number of adherent and emigrated leukocytes
in both diabetic and nondiabetic rats, but leukocyte adhesion and
emigration after reperfusion were significantly higher in diabetic than
control rats (Fig 2
). Albumin leakage increased significantly
in response to ischemia-reperfusion in both groups of rats;
however, diabetic rats exhibited a significantly higher albumin
leakage than control rats after reperfusion (Fig 3).
|
) and control (
) rats.
+P<.05 vs basal; *P<.05 vs
controls.In control rats, superfusion of the mesentery with bicarbonate buffer containing 480 mg/dL glucose did not produce any significant change in red blood cell velocity, shear rate, leukocyte rolling, adhesion, emigration, or microvascular permeability (data not shown). In this series of experiments, the responses of leukocyte rolling (0.28±0.06 cells/100 µm), adherence (6.66±0.88 cells/100 µm), and emigration (5.00±0.57 cells per field) to ischemia-reperfusion and the increased albumin leakage (0.24±0.05) were very similar to those observed in control preparations and significantly different from the response observed in the diabetic group.
Infusion of 50% glucose at a rate that increased plasma glucose concentration to diabetic levels (498±21 and 534±33 mg/dL IV at 30 and 80 minutes of continuous infusion, respectively) produced a rapid and sustained increase in red blood cell velocity (3.26±0.25 versus 3.72±0.28 mm/s, P<.01), an increase in shear rate (556±22 versus 634±24 seconds, P<.01), and a decrease in the number of rolling leukocytes (0.59±13 versus 0.29±.06 cells/100 µm, P<.01) without significant changes in leukocyte adhesion (2.2±0.58 versus 2.4±0.51 cells/100 µm), emigration (1.81±0.58 versus 1.99±0.68 cells per field), or albumin leakage (0.077±0.024 versus 0.076±0.019) compared with baseline values. In these experiments, the ischemia-reperfusion–induced changes in leukocyte rolling (0.31±0.10 cells/100 µm), adherence (4.60±1.20 cells/100 µm), and emigration (2.80±0.49 cells per field) and the increased albumin leakage (0.29±0.06) were very similar to responses observed in the control group and significantly different from those noted in the diabetic group.
In diabetic rats, administration of a P-selectin mAb had a marked
effect on the pattern of leukocyte rolling observed 5 minutes after
reperfusion. The P-selectin mAb resulted in a significant increase in
leukocyte rolling velocity and reduced both the flux and number of
rolling leukocytes compared with untreated diabetic rats and those
receiving a nonbinding mAb (Fig 5). At 30 minutes after
reperfusion, rats treated with the P-selectin mAb still exhibited a
significantly higher leukocyte rolling velocity compared with baseline
values, but differences among groups were not significant. P-selectin
mAb treatment also was associated with a significant reduction in the
number of adherent (Fig 6A) and emigrated (Fig
6B)
leukocytes. Treatment with an L-selectin mAb had less
effect on leukocyte rolling. The L-selectin mAb prevented
the reduction in leukocyte rolling velocity and the increases in the
flux and number of rolling leukocytes that usually were observed 5
minutes after reperfusion in untreated rats and rats treated with a
nonbinding mAb (Fig 5). Although there was a trend for reduced
ischemia-reperfusion–induced leukocyte adhesion and
emigration in rats treated with the L-selectin mAb
(relative to untreated rats and those receiving a nonbinding mAb),
these differences were not statistically significant. Administration of
mAbs directed against either CD11/CD18 or ICAM-1 had no effect on
leukocyte rolling at reperfusion (data not shown) but markedly
decreased leukocyte adhesion and emigration after reperfusion (Fig
6).
In diabetic rats, none of the mAbs studied exerted a significant effect
on albumin leakage after ischemia-reperfusion (Fig 7).
|
, treated with P-selectin mAb.
+P<.05 vs basal; *P<.05 vs
controls.
|
|
Baseline and ischemia-reperfusion values for centerline red blood cell velocity and shear rate in postcapillary venules of diabetic rats were not altered by treatment with any of the mAbs (binding or nonbinding).
|
Discussion |
|---|
|
TopAbstractIntroductionMethods Results Discussion References |
|---|
The results of this study indicate that the diabetic state induces an exaggerated inflammatory response to ischemia-reperfusion manifested as a greater accumulation of adherent and emigrated leukocytes and a larger increase in albumin extravasation. The present study also shows that diabetes is associated with an increased baseline level of leukocyte rolling (Fig 1
). Although we cannot exclude the possibility of
altered
adhesion molecule expression in diabetic rats as a basis for the
observed changes in leukocyte rolling, a more likely explanation for
this response is the corresponding reduction in venular blood flow
noted in mesenteric venules of diabetic rats. Previous
studies24 25 have demonstrated a fairly constant
ratio of
leukocyte rolling velocity to red cell velocity over a wide range of
shear rates in mesenteric venules, and our finding that this
ratio does not differ between control and diabetic rats
suggests that changes in blood flow velocity account largely for
reduced leukocyte rolling velocity in diabetic venules. The decreased
red blood cell velocity and shear rates observed in diabetic rats might
be related to a redistribution of blood flow through the net of
proliferating small vessels that we observed in the mesentery 3 weeks
after the induction of diabetes.
Despite the higher basal number of rolling leukocytes in diabetic rats
relative to control rats, the former group also exhibited a more marked
recruitment of rolling leukocytes in response to
ischemia-reperfusion (Fig 4). In control rats,
ischemia-reperfusion induced a transient increase in the
number of rolling leukocytes that was determined primarily by an
increased flux of rolling leukocytes without significant changes in
leukocyte rolling velocity. However, the increased number of rolling
leukocytes observed in diabetic rats after
ischemia-reperfusion resulted from both an increased flux
of rolling leukocytes and a further decrease in leukocyte rolling
velocity. The latter observation suggests that the additional increase
in rolling leukocytes in diabetic rats may result from a shear
rate–dependent recruitment of rollers.25
Our study also provides some insights into the contributions of
leukocyte (L-selectin, CD11/CD18) and
endothelial cell (P-selectin, ICAM-1) adhesion
molecules to the ischemia-reperfusion–induced
recruitment of rolling leukocytes in diabetic rats (Fig 5). We
noted
that mAbs directed against either P-selectin or L-selectin
were effective in blunting the leukocyte rolling response elicited by
ischemia-reperfusion, whereas mAbs against CD11/CD18 or
ICAM-1 and a nonbinding (control) mAb had no effect. These observations
suggest that the altered leukocyte rolling response elicited by
ischemia-reperfusion is mediated primarily by adhesive
interactions of L-selectin on leukocytes and P-selectin on
endothelial cells. The involvement of P-selectin in
mediating the rapid rolling response elicited by
ischemia-reperfusion is consistent with the
rapidity of expression of the lectinlike adhesion molecule after
activation of endothelial cells in
culture26 and the well-characterized role of
P-selectin in mediating leukocyte rolling in mesenteric venules exposed
to normal27 28 and low25 shear rates.
Our
inability to demonstrate a contribution of CD11/CD18 and ICAM-1 to the
enhanced leukocyte rolling in diabetes is also in accordance with
published reports that failed to invoke a role for these adhesion
molecules in the modulation of leukocyte
rolling.23 29
Kurose and associates10 recently demonstrated that rat mesenteric venules exposed to ischemia-reperfusion sustain an increased number of firmly adherent and emigrating leukocytes. Our findings in control rats confirm their observations, and our results in diabetic rats extend their work to show that the leukocyte–endothelial cell adhesive interactions that are manifested as adherence and emigration in postischemic microvessels are greatly exaggerated in this animal model of human disease. The quantitatively different responses to ischemia-reperfusion between control and diabetic rats are not due to differential ischemic insults inasmuch as venular red blood cell velocity during ischemia fell to a similar level in both groups of rats. However, a lower venular shear rate or a higher number of rolling leukocytes in diabetic venules after reperfusion could account for at least some of the differences in leukocyte adherence and emigration between control and diabetic rats. A dependency of firm leukocyte adhesion on shear rates has been demonstrated in vitro30 and in vivo.24 It also has been shown that graded reductions in rolling leukocyte flux are accompanied by proportional reductions in chemoattractant-induced leukocyte adhesion.31 Taken together, these observations suggest that the lower shear rates in diabetic compared with control rats may contribute to the increased leukocyte adhesion observed after ischemia-reperfusion in diabetic rats. Nonetheless, we cannot invoke differences in shear rate as the sole explanation for the exaggerated inflammatory response to ischemia-reperfusion in diabetic rats, particularly because these differences in shear rate were already present under baseline conditions, when similar numbers of adherent leukocytes were observed in control and diabetic rats.
The present study also demonstrates that hyperglycemia per se cannot explain the altered inflammatory responses observed in diabetic rats. Elevation of blood glucose concentration in control rats to a level measured in diabetic rats induced significant increases in red blood cell velocity and venular shear rate and a decrease in the number of rolling leukocytes relative to baseline conditions. These changes contrast the microvascular alterations noted in diabetic rats. In addition, acute hyperglycemia did not enhance the inflammatory responses (leukocyte–endothelial cell adhesion and albumin leakage) usually elicited by ischemia-reperfusion.
In postcapillary venules of control (nondiabetic) rats subjected to
ischemia-reperfusion32 33 34 and on
endothelial cell monolayers exposed to anoxia or
reoxygenation,35 the increased
leukocyte adherence is mediated by an interaction between CD11/CD18 on
leukocytes and ICAM-1 on endothelial cells. A recent
study from our group10 characterized the molecular
determinants of ischemia-reperfusion–induced leukocyte
adhesion and emigration in nondiabetic rats. That study showed that
mAbs directed against CD18 and ICAM-1 but not P- or
L-selectin effectively blunted
ischemia-reperfusion–induced leukocyte adherence and
emigration in rat mesenteric venules; these results are summarized in
the Table for comparison with the current findings in
diabetic rats. In the present study, we provide evidence that
CD11/CD18–ICAM interactions also are important in mediating the
leukocyte adherence and emigration observed in mesenteric venules of
diabetic rats exposed to ischemia-reperfusion (Fig 6). However,
a more substantial role for selectins as a determinant of
ischemia-reperfusion–induced leukocyte adherence can
be inferred from our experiments on diabetic rats. The P-selectin mAb,
which had a very potent and lasting effect on leukocyte rolling after
ischemia-reperfusion in diabetic rats, effectively reduced
both leukocyte adherence and emigration. The L-selectin
mAb, which had a less potent and short-lived effect on leukocyte
rolling, was less effective in reducing leukocyte adherence and had no
significant effect on leukocyte emigration. These observations suggest
that a very effective blockade of leukocyte rolling is needed to
completely prevent firm adhesion. An explanation for the relatively
striking effect of the P-selectin mAb in reducing leukocyte adherence
and emigration in diabetic rats (compared with published studies on
control rats) is not readily available; however, it may reflect
differences in the ability of ischemia-reperfusion to
mobilize P-selectin to the endothelial cell surface
between control and diabetic rats.
|
Numerous reports demonstrate an increased microvascular permeability to
plasma proteins in tissues exposed to ischemia-reperfusion
and show that the ischemia-reperfusion–induced
vascular protein leakage is mediated by adherent
leukocytes.32 36 37 38 Thus,
another primary objective of
this study was to determine whether diabetes alters the phenomenon of
ischemia-reperfusion–induced
endothelial barrier dysfunction. Our results indicate
that even under basal conditions, albumin leakage from
mesenteric venules is significantly higher in diabetic compared with
control rats (Fig 3). This observation is consistent with
published evidence demonstrating an increased microvascular
permeability in different organs of diabetic
rats.39 40
However, despite this basal level of endothelial cell
barrier dysfunction, diabetic rats exhibited a significantly enhanced
albumin leakage in response to
ischemia-reperfusion. However, unlike control rats in which
mAbs that attenuate ischemia-reperfusion–induced
leukocyte–endothelial cell adhesion also
blunt the accompanying albumin leakage,10
postcapillary venules of diabetic rats are largely unresponsive to
immunoneutralization of adhesion molecules on leukocytes or
endothelial cells (see the Table). These observations
indicate that the enhanced albumin extravasation observed in
control and postischemic venules of diabetic rats is
mediated by leukocyte-independent processes.
It has been proposed that hyperglycemia per se may contribute to the impaired endothelial cell barrier function in diabetes.41 However, the findings of the present study do not favor this hypothesis; we observed that exposure of the mesenteric microvasculature to high glucose levels did not alter albumin leakage, either under baseline conditions or after ischemia-reperfusion. Other leukocyte-independent mechanisms that may contribute to the enhanced microvascular protein transfer in diabetes include changes in redox state in endothelial cells as a result of glucose metabolism through the polyol pathway,42 activation of protein kinase C,43 and quenching of nitric oxide by advanced glycosylation products.44 45
|
Acknowledgments |
|---|
This work was supported by NIH grants HL-26441 and DK-32288. Dr Panés is a recipient of a grant from Fundación Ramón Areces.
Received April 25, 1995; revision received August 16, 1995; accepted August 20, 1995.
|
References |
|---|
|
TopAbstractIntroductionMethods Results Discussion References |
|---|
1. Ganda OP. Pathogenesis of macrovascular disease in the human diabetic. Diabetes. 1980;29:931-942. [Medline] [Order article via Infotrieve]
2. Jensen T, Feldt-Rasmussen B, Bjerre-Knudsen J, Deckert T. Features of endothelial dysfunction in early diabetic nephropathy. Lancet. 1989;2:461-463. [Medline] [Order article via Infotrieve]
3. Suzuki H, Zweifach BW, Forrest MJ, Schmid-Schonbein GW. Modification of leukocyte adhesion in spontaneously hypertensive rats by adrenal corticosteroids. J Leukoc Biol. 1995;57:20-26. [Abstract]
4. Lehr HA, Arfors KE, Hübner C, Menger MD, Messmer K. Leukocyte-endothelial cell interaction as a target for antiatherogenic strategies in allograft transplantation. Transplant Proc. 1993;25:2067-2069. [Medline] [Order article via Infotrieve]
5. del Zoppo GJ, García JH. Polymorphonuclear leukocyte adhesion in cerebrovascular ischemia. In: Granger DN, Schmid-Schönbein GW, eds. Physiology and Pathophysiology of Leukocyte Adhesion. New York, NY: Oxford University Press; 1995:408-433.
6. Lefer DJ. Role of leukocyte adhesion in myocardial ischemia-reperfusion injury. In: Granger DN, Schmid-Schönbein GW, eds. Physiology and Pathophysiology of Leukocyte Adhesion. New York, NY: Oxford University Press; 1995:393-407.
7.
Korthuis RJ, Grisham MB, Zimmerman BJ, Granger DN,
Taylor AE. Vascular injury in dogs during
ischemia-reperfusion: improvement with
ATP-MgCl2 pretreatment. Am J Physiol. 1988;254:H702-H708.
8.
Kraemer R, Smith CW, Mullane KM.
Activated human polymorphonuclear leucocytes reduce rabbit
papillary muscle function: role of the CD18 glycoprotein
adhesion complex. Cardiovasc Res. 1991;25:172-175.
9. Wallace JL, Arfors KE, McKnight GW. A monoclonal antibody against the CD18 leukocyte adhesion molecule prevents indomethacin-induced gastric damage in the rabbit. Gastroenterology. 1991;100:878-883. [Medline] [Order article via Infotrieve]
10.
Kurose I, Anderson DC, Miyasaka M, Tamatani T, Paulson
JC, Todd RF, Rusche JR, Granger DN. Molecular determinants of
reperfusion-induced leukocyte adhesion and vascular protein
leakage. Circ Res. 1994;74:336-343.
11. Granger DN, Kurose I, Kvietys PR. Modulation of leukocyte adherence and emigration during ischemia and reperfusion. In: Granger DN, Schmid-Schönbein GW, eds. Physiology and Pathophysiology of Leukocyte Adhesion. New York, NY: Oxford University Press; 1995:323-338.
12. Kelly LW, Barden CA, Tiedeman JS, Hatchell DL. Alterations in viscosity and filterability of whole blood and blood cell subpopulations in diabetic cats. Exp Eye Res. 1993;56:341-347. [Medline] [Order article via Infotrieve]
13. Fisher TC, Meiselmann HJ. Polymorphonuclear leukocytes in ischemic vascular disease. Thromb Res. 1994;74(suppl 1):S21-S34.
14. Masuda M, Murakami T, Egawa H, Murata K. Decreased fluidity of polymorphonuclear leukocyte membrane in streptozocin-induced diabetic rats. Diabetes. 1990;39:466-470. [Abstract]
15. Freedman SF, Hatchell DL. Enhanced superoxide radical production by stimulated polymorphonuclear leukocytes in a cat model of diabetes. Exp Eye Res. 1992;55:767-773. [Medline] [Order article via Infotrieve]
16. Wierusz-Wysocka B, Wysocki H, Siekierka H, Wykretowicz A, Szczepanik A, Klimas R. Evidence of polymorphonuclear neutrophils (PMN) activation in patients with insulin-dependent diabetes mellitus. J Leukoc Biol. 1987;42:519-523. [Abstract]
17. Bohlen HG, Hankins KD. Early arteriolar and capillary changes in streptozotocin-induced diabetic rats and intraperitoneal hyperglycaemic rats. Diabetologia. 1982;22:344-348. [Medline] [Order article via Infotrieve]
18. Mulligan MS, Varani J, Warren JS, Till GO, Smith CW, Anderson DC, Todd RD, Ward PA. Roles of beta 2 integrins of rat neutrophils in complement- and oxygen radical-mediated acute inflammatory injury. J Immunol. 1992;148:1847-1857. [Abstract]
19. Tamatani T, Miyasaka M. Identification of monoclonal antibodies reactive with the rat homolog of ICAM-1, and evidence for a differential involvement of ICAM-1 in the adherence of resting versus activated lymphocytes to high endothelial cells. Int J Immunol. 1990;2:165-171.
20. Tamatani T, Kitamura F, Kuida K, Shirao M, Mochizuki M, Suematsu M, Schmid-Schönbein GW, Watanabe K, Tsurufuji S, Miyasaka M. Characterization of rat LECAM-1 (L-selectin) by the use of monoclonal antibodies and evidence for the presence of soluble LECAM-1 in rat sera. Eur J Immunol. 1993;23:2181-2188. [Medline] [Order article via Infotrieve]
21. Mulligan MS, Polley MJ, Bayer RJ, Nunn MF, Paulson JC, Ward PA. Neutrophil-dependent acute lung injury: requirement for P-selectin (GMP-140). J Clin Invest. 1992;90:1600-1607.
22. Winn RK, Liggitt D, Vedder NB, Paulson JC, Harlan JM. Anti-P-selectin monoclonal antibody attenuates reperfusion injury to the rabbit ear. J Clin Invest. 1993;92:2042-2047.
23.
Zimmerman BJ, Holt JW, Paulson JC, Anderson DC,
Miyasaka M, Tamatani T, Todd RF, Rusche JR, Granger DN.
Molecular determinants of lipid mediator-induced leukocyte
adherence and emigration in rat mesenteric venules. Am J
Physiol. 1994;266:H847-H853.
24. Perry MA, Granger DN. Role of CD11/CD18 in shear rate-dependent leukocyte-endothelial cell interactions in cat mesenteric venules. J Clin Invest. 1991;87:1798-1804.
25.
Bienvenu K, Russell J, Granger DN.
Leukotriene B4 mediates shear rate–dependent leukocyte
adhesion in mesenteric venules. Circ Res. 1992;71:906-911.
26. McEver RP. GMP-140: a receptor for neutrophils and monocytes on activated platelets and endothelium. J Cell Biochem. 1991;45:156-161. [Medline] [Order article via Infotrieve]
27.
Dore M, Korthuis RJ, Granger DN, Entman ML, Smith
CW. P-selectin mediates spontaneous leukocyte rolling in
vivo. Blood. 1993;82:1308-1316.
28.
Gaboury JP, Anderson DC, Kubes P. Molecular
mechanisms involved in superoxide-induced
leukocyte-endothelial cell interactions in
vivo. Am J Physiol. 1994;266:H637-H642.
29.
Bienvenu K, Granger DN. Molecular determinants
of shear rate-dependent leukocyte adhesion in postcapillary
venules. Am J Physiol. 1993;264:H1504-H1508.
30.
Ley K, Lundgren E, Berger E, Arfors KE.
Shear-dependent inhibition of granulocyte adhesion to cultured
endothelium by dextran sulfate.
Blood. 1989;73:1324-1330.
31. Lindbom L, Xie X, Raud J, Hedqvist P. Chemoattractant-induced firm adhesion of leukocytes to vascular endothelium in vivo is critically dependent on initial leukocyte rolling. Acta Physiol Scand. 1992;146:415-421. [Medline] [Order article via Infotrieve]
32.
Hernandez LA, Grisham MB, Twohig B, Arfors KE, Harlan
JM, Granger DN. Role of neutrophils in
ischemia-reperfusion-induced microvascular
injury. Am J Physiol. 1987;253:H699-H703.
33.
Kurtel H, Tso P, Granger DN. Granulocyte
accumulation in postischemic intestine: role of leukocyte
adhesion glycoprotein CD11/CD18. Am J
Physiol. 1992;262:G878-G882.
34. Suzuki M, Asako H, Kubes P, Jennings S, Grisham MB, Granger DN. Neutrophil-derived oxidants promote leukocyte adherence in postcapillary venules. Microvasc Res. 1991;42:125-138. [Medline] [Order article via Infotrieve]
35. Yoshida N, Granger DN, Anderson DC, Rothlein R, Lane C, Kvietys PR. Anoxia/reoxygenation-induced neutrophil adherence to cultured endothelial cells. Am J Physiol. 1992;292:H1891-H1898.
36.
Grisham MB, Hernandez LA, Granger DN.
Adenosine inhibits ischemia-reperfusion-induced
leukocyte adherence and extravasation. Am J Physiol. 1989;257:H1334-H1339.
37.
Adkins WK, Taylor AE. Role of xanthine oxidase
and neutrophils in ischemia-reperfusion injury in rabbit
lung. J Appl Physiol. 1990;69:2012-2018.
38.
Korthuis RJ, Grisham MB, Granger DN. Leukocyte
depletion attenuates vascular injury in postischemic
skeletal muscle. Am J Physiol. 1988;254:H823-H827.
39.
Korthuis RJ, Pitts VH, Granger DN. Intestinal
capillary filtration in experimental diabetes mellitus.
Am J Physiol. 1987;253:G20-G25.
40. Huijberts MS, Wolffenbuttel BH, Crijns FR, Nieuwenhuijzen-Kruseman AC, Bemelmans MH, Struijker-Boudier HA. Aminoguanidine reduces regional albumin clearance but not urinary albumin excretion in streptozotocin-diabetic rats. Diabetologia. 1994;37:10-14. [Medline] [Order article via Infotrieve]
41. Ruderman NB, Williamson JR, Brownlee M. Glucose and diabetic vascular disease. FASEB J. 1992;6:2905-2914. [Abstract]
42. Tilton RG, Chang K, Pugliese G, Eades DM, Province MA, Sherman WR, Kilo C, Williamson JR. Prevention of hemodynamic and vascular albumin filtration changes in diabetic rats by aldose reductase inhibitors. Diabetes. 1989;38:1258-1270. [Abstract]
43. Lynch JJ, Ferro TJ, Blumenstock FA, Brockenauer AM, Malik AB. Increased endothelial albumin permeability mediated by protein kinase C activation. J Clin Invest. 1990;85:1991-1998.
44. Bucala R, Tracey KJ, Cerami A. Advanced glycosylation products quench nitric oxide and mediate defective endothelium-dependent vasodilatation in experimental diabetes. J Clin Invest. 1991;87:432-438.
45.
Kubes P, Granger DN. Nitric oxide modulates
microvascular permeability. Am J Physiol. 1992;262:H611-H615.
This article has been cited by other articles:
 |
|
 |
|
  G.-Y. Song, Y.-J. Wu, Y.-J. Yang, J.-J. Li, H.-L. Zhang, H.-J. Pei, Z.-Y. Zhao, Z.-H. Zeng, and R.-T. Hui The accelerated post-infarction progression of cardiac remodelling is associated with genetic changes in an untreated streptozotocin-induced diabetic rat model Eur J Heart Fail, October 1, 2009; 11(10): 911 - 921. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. N. Vasdekis, M. Argentou, J. D. Kakisis, A. Bossios, D. Gourgiotis, M. Karanikolas, and G. Karatzas A Global Assessment of the Inflammatory Response Elicited Upon Open Abdominal Aortic Aneurysm Repair Vascular and Endovascular Surgery, March 1, 2008; 42(1): 47 - 53. [Abstract] [PDF]
|
|
|
|
 |
|
 R. Scalia, Y. Gong, B. Berzins, L. J. Zhao, and K. Sharma Hyperglycemia Is a Major Determinant of Albumin Permeability in Diabetic Microcirculation: The Role of {micro}-Calpain Diabetes, July 1, 2007; 56(7): 1842 - 1849. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Shankar, R. Klein, B. E.K. Klein, F. J. Nieto, and S. E. Moss Relationship Between Low-Normal Blood Pressure and Kidney Disease in Type 1 Diabetes Hypertension, January 1, 2007; 49(1): 48 - 54. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Martin, S. Rojas, A. Chamorro, C. Falcon, N. Bargallo, and A. M. Planas Why Does Acute Hyperglycemia Worsen the Outcome of Transient Focal Cerebral Ischemia?: Role of Corticosteroids, Inflammation, and Protein O-Glycosylation Stroke, May 1, 2006; 37(5): 1288 - 1295. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. J. M. Greer, D. P. Ware, and D. J. Lefer Myocardial infarction and heart failure in the db/db diabetic mouse Am J Physiol Heart Circ Physiol, January 1, 2006; 290(1): H146 - H153. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. Bito, S. Hino, A. Baba, M. Tanaka, H. Watabe, and H. Kawabata Degradation of oxidative stress-induced denatured albumin in rat liver endothelial cells Am J Physiol Cell Physiol, September 1, 2005; 289(3): C531 - C542. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. J. Stalker, Y. Gong, and R. Scalia The Calcium-Dependent Protease Calpain Causes Endothelial Dysfunction in Type 2 Diabetes Diabetes, April 1, 2005; 54(4): 1132 - 1140. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. M. Yada-Langui, E. A. Anjos-Valotta, P. Sannomiya, M. Rocha e Silva, and R. Coimbra Resuscitation Affects Microcirculatory Polymorphonuclear Leukocyte Behavior After Hemorrhagic Shock: Role of Hypertonic Saline and Pentoxifylline Experimental Biology and Medicine, July 1, 2004; 229(7): 684 - 693. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. N. Granger, T. Vowinkel, and T. Petnehazy Modulation of the Inflammatory Response in Cardiovascular Disease Hypertension, May 1, 2004; 43(5): 924 - 931. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Galinanes and A. G Fowler Role of clinical pathologies in myocardial injury following ischaemia and reperfusion Cardiovasc Res, February 15, 2004; 61(3): 512 - 521. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Canbaz, E. Duran, and A. Lassnigg Ischaemia-reperfusion studies and diabetes mellitus Br. J. Anaesth., July 1, 2003; 91(1): 158 - 159. [Full Text] [PDF]
|
|
|
|
 |
|
 J. Melin, O. Hellberg, and B. Fellstrom Hyperglycaemia and renal ischaemia-reperfusion injury Nephrol. Dial. Transplant., March 1, 2003; 18(3): 460 - 462. [Full Text] [PDF]
|
|
|
|
|
|
P. Algenstaedt, C. Schaefer, T. Biermann, A. Hamann, B. Schwarzloh, H. Greten, W. Ruther, and N. Hansen-Algenstaedt Microvascular Alterations in Diabetic Mice Correlate With Level of Hyperglycemia Diabetes, February 1, 2003; 52(2): 542 - 549. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Sola, J. Panes, C. Xaus, and G. Hotter Fructose-1,6-biphosphate and nucleoside pool modifications prevent neutrophil accumulation in the reperfused intestine J. Leukoc. Biol., January 1, 2003; 73(1): 74 - 81. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H. Vlassara, W. Cai, J. Crandall, T. Goldberg, R. Oberstein, V. Dardaine, M. Peppa, and E. J. Rayfield Inflammatory mediators are induced by dietary glycotoxins, a major risk factor for diabetic angiopathy PNAS, November 26, 2002; 99(24): 15596 - 15601. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. Booth, T. J. Stalker, A. M. Lefer, and R. Scalia Mechanisms of Amelioration of Glucose-Induced Endothelial Dysfunction Following Inhibition of Protein Kinase C In Vivo Diabetes, May 1, 2002; 51(5): 1556 - 1564. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. C. Thomas, T. H. Mathew, and G. R. Russ Glycaemic control and graft loss following renal transplantation Nephrol. Dial. Transplant., October 1, 2001; 16(10): 1978 - 1982. [Full Text] [PDF]
|
|
|
|
 |
|
 G. Booth, T. J. Stalker, A. M. Lefer, and R. Scalia Elevated ambient glucose induces acute inflammatory events in the microvasculature: effects of insulin Am J Physiol Endocrinol Metab, June 1, 2001; 280(6): E848 - E856. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. Boeri, M. Maiello, and M. Lorenzi Increased Prevalence of Microthromboses in Retinal Capillaries of Diabetic Individuals Diabetes, June 1, 2001; 50(6): 1432 - 1439. [Abstract] [Full Text]
|
|
|
|
 |
|
 L. Fontana, C. Giagulli, P. Minuz, A. Lechi, and C. Laudanna 8-Iso-PGF2{{alpha}} Induces {beta}2-Integrin-Mediated Rapid Adhesion of Human Polymorphonuclear Neutrophils : A Link Between Oxidative Stress and Ischemia/Reperfusion Injury Arterioscler Thromb Vasc Biol, January 1, 2001; 21(1): 55 - 60. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Tsujikawa, J. Kiryu, A. Nonaka, K. Yamashiro, H. Nishiwaki, Y. Honda, and Y. Ogura Leukocyte-endothelial cell interactions in diabetic retina after transient retinal ischemia Am J Physiol Regulatory Integrative Comp Physiol, September 1, 2000; 279(3): R980 - R989. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Hoshida, N. Yamashita, K. Otsu, T. Kuzuya, and M. Hori Cholesterol feeding exacerbates myocardial injury in Zucker diabetic fatty rats Am J Physiol Heart Circ Physiol, January 1, 2000; 278(1): H256 - H262. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Chello, P. Mastroroberto, F. Cirillo, E. Bevacqua, A. Carrano, F. Perticone, and A. R. Marchese Neutrophil-endothelial cells modulation in diabetic patients undergoing coronary artery bypass grafting Eur. J. Cardiothorac. Surg., October 1, 1999; 14(4): 373 - 379. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. L. Martinez, M. Aparecida de Oliveira, and Z. B. Fortes Influence of Verapamil and Diclofenac on Leukocyte Migration in Rats Hypertension, October 1, 1999; 34(4): 997 - 1001. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. J. Korthuis, D. C. Gute, F. Blecha, and C. R. Ross PR-39, a proline/arginine-rich antimicrobial peptide, prevents postischemic microvascular dysfunction Am J Physiol Heart Circ Physiol, September 1, 1999; 277(3): H1007 - H1013. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
I. Kurose, R. Wolf, W. Cerwinka, and D. N. Granger Microvascular Responses to Ischemia/Reperfusion in Normotensive and Hypertensive Rats Hypertension, August 1, 1999; 34(2): 212 - 216. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. R. Kersten and D. C. Warltier Modulation of the adaptive response to myocardial ischemia by coexisting disease Am J Physiol Heart Circ Physiol, June 1, 1999; 276(6): H2268 - H2270. [Full Text] [PDF]
|
|
|
|
|
|
N. Mori, Y. Horie, M. E. Gerritsen, and D. N. Granger Ischemia-reperfusion induced microvascular responses in LDL-receptor -/- mice Am J Physiol Heart Circ Physiol, May 1, 1999; 276(5): H1647 - H1654. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Salas, J. Panes, J. I. Elizalde, M. Casadevall, D. C. Anderson, D. N. Granger, and J. M. Pique Mechanisms responsible for enhanced inflammatory response to ischemia-reperfusion in diabetes Am J Physiol Heart Circ Physiol, November 1, 1998; 275(5): H1773 - H1781. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. S. Tsao, J. Niebauer, R. Buitrago, P. S. Lin, B.-y. Wang, J. P. Cooke, Y-d. Ida Chen, and G. M. Reaven Interaction of Diabetes and Hypertension on Determinants of Endothelial Adhesiveness Arterioscler Thromb Vasc Biol, June 1, 1998; 18(6): 947 - 953. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Komatsu, J. Panes, J. M. Russell, D. C. Anderson, V. R. Muzykantov, M. Miyasaka, and D. N. Granger Effects of Chronic Arterial Hypertension on Constitutive and Induced Intercellular Adhesion Molecule-1 Expression In Vivo Hypertension, February 1, 1997; 29(2): 683 - 689. [Abstract] [Full Text]
|
|
| |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||