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1 Department of Physiology, Faculty of Medicine, Akdeniz University, Antalya, 07070 Turkey; and 2 Department of Physiology and Biophysics, Keck School of Medicine, Los Angeles, California 90033
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ABSTRACT |
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 TOP
 ABSTRACT
 INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
REFERENCES
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In addition to
its known action on vascular smooth muscle, nitric oxide (NO) has been
suggested to have cardiovascular effects via regulation of red blood
cell (RBC) deformability. The present study was designed to further
explore this possibility. Human RBCs in autologous plasma were
incubated for 1 h with NO synthase (NOS) inhibitors
[N
-nitro-L-arginine methyl ester
(L-NAME) and S-methylisothiourea], NO donors
[sodium nitroprusside (SNP) and diethylenetriamine
(DETA)-NONOate], an NO precursor (L-arginine), soluble
guanylate cyclase inhibitors (1H-[1,2,4]oxadiazolo-[4,3-a]quinoxalin-1-one and methylene
blue), and a potassium channel blocker [triethylammonium (TEA)].
After incubation, RBC deformability at various shear stresses was
determined by ektacytometry. Both NOS inhibitors significantly reduced
RBC deformability above a threshold concentration, whereas the NO donors increased deformability at optimal concentrations. NO donors, as
well as the NO precursor L-arginine and the potassium
blocker TEA, were able to reverse the effects of NOS inhibitors.
Guanylate cyclase inhibition reduced RBC deformation, with both SNP and DETA-NONOate able to reverse this effect. These results thus indicate the importance of NO as a determinant of RBC mechanical behavior and
suggest its regulatory role for normal RBC deformability.
hemorheology; hemodynamics; regulation; cGMP
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INTRODUCTION |
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TOP
ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
REFERENCES
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NITRIC OXIDE (NO) plays a major role in cardiovascular regulation (5, 27), with its action mainly attributed to the effects on vascular smooth muscle cells (5, 25, 27). However, it has also been shown that NO synthesized in endothelial cells not only diffuses to the adjacent smooth muscle cells but also to the vascular lumen (7, 24). In addition to its effects on leukocytes and platelets, NO interacts with red blood cells (RBCs) via binding to the heme portion of hemoglobin to form S-nitrosohemoglobin (16, 45) and inducing the formation of methemoglobin (23, 43, 45).
Human RBCs are positive for both inducible (NOS2 or iNOS) and constitutive (NOS3 or eNOS) forms of NO synthase (NOS), and thus are capable of synthesizing their own NO (17). Petrov et al. (37, 38) have demonstrated the existence of particulate and soluble guanylate cyclase as well as phosphodiesterases in human RBCs. It has therefore been suggested that NO synthesized within RBCs may modulate RBC physiological behavior and thus that both extracellular and intracellular sources of NO can affect the cell (9, 17).
Korbut and co-workers (21, 22, 41) have suggested that NO
may have a regulatory effect on RBC deformability and aggregation and
have shown that this effect is concentration dependent. The same
authors also demonstrated that NOS inhibitors have a protective effect
on erythrocyte deformability in septic shock and in the acute phase of
endotoxemia in rats (22, 42). Furthermore, Mesquita et al.
(29) have shown that both acetylcholine (ACh) and the NO
donor spermine-NONOate improve RBC deformability and suggest that the
effect of ACh may be due to induced NO synthesis mediated by
M1-type ACh receptors on the RBC membrane. Chronic inhibition of NOS by
N-nitro-L-arginine methyl ester
(L-NAME) has been found to significantly reduce RBC
deformability in rats (6); the mechanical properties of
RBC from these animals were normalized in vitro by a low dose (10 µM)
of sodium nitroprusside (SNP), whereas higher doses were ineffective
(6).
The present study was designed to provide further insight into the effects of NO on the rheological behavior of human RBCs. In particular, the in vitro effects of NOS inhibitors, NO donors, and guanylate cyclase inhibitors on RBC deformation in defined shear fields were evaluated. Our results indicate the marked influence of NO on RBC deformability and thus lend support to the possibility that NO has an important regulatory effect on this cellular mechanical property.
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MATERIALS AND METHODS |
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TOP
ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
REFERENCES
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Blood Sampling
Blood samples, anticoagulated with heparin (15 IU/ml), were obtained from healthy adult male volunteers. RBCs were isolated from whole blood by centrifugation (1,400 g, 6 min) followed by two washing steps in PBS (pH = 7.4, 290 mosmol/kg). The washed RBCs were then resuspended in plasma at a hematocrit of 40% for the experimental studies. All RBC preparations, incubations, and measurements were carried out within 4-6 h after blood collection.Incubations
RBC suspensions were incubated at room temperature for 1 h in the presence of various chemical agents, after which RBCs from these suspensions were used for the determination of RBC size, geometry, and cellular deformability. This 60-min incubation period was selected based on preliminary studies that indicated a gradual increase of effects up to 40 min, with no additional change thereafter. The following agents were used for the incubations.NOS inhibitors and/or NO donors.
Both a nonspecific NOS inhibitor (L-NAME) and a specific
iNOS inhibitor [S-methylisothiourea (SMT)] were used.
The effects of these NOS inhibitors were first tested at concentrations
between 10
6 and 102 M, and the most
effective doses were then selected to be used for the rest of the
L-NAME and SMT studies. SNP and diethylenetriamine (DETA)-NONOate were used as NO donors at concentrations between 104 and 107 M and were employed alone or
with NOS inhibitors.
NO precursor.
The effect of the NO precursor L-arginine was evaluated at
a concentration of 105 M.
Soluble guanylate cyclase inhibitors.
The soluble guanylate cyclase inhibitor
1H-[1,2,4]oxadiazolo-[4,3-a]quinoxalin-1-one (ODQ) was
used with or without NO donors. ODQ was tested at various doses
(106-104 M), after which
105 M was selected to be used throughout the study.
Additionally, in a separate series of experiments, methylene blue, an
inhibitor of both NOS and soluble guanylate cyclase, was tested at
105 M.
Potassium channel inhibitor.
The effect of the nonselective potassium channel inhibitor
tetraethylammonium chloride (TEA) was first tested at concentrations between 104 and 1010 M and then used at
108 M with or without L-NAME.
RBC Deformability Measurements
RBC deformability (i.e., the ability of the entire cell to adopt a new configuration when subjected to applied mechanical forces) was determined at various fluid shear stresses by laser diffraction analysis using an ektacytometer (LORCA, RR Mechatronics; Hoorn, The Netherlands). The system has been described elsewhere in detail (18). Briefly, a low hematocrit suspension of RBC in an isotonic viscous medium (70-kDa dextran) was sheared in a Couette system composed of a glass cup and a precisely fitting bob, with a gap of 0.3 mm between the cylinders. A laser beam was directed through the sheared sample, and the diffraction pattern produced by the deformed cells was analyzed by a microcomputer. On the basis of the geomerty of the ellipitical diffraction pattern, an elongation index (EI) was calculated: EI = (L
W)/(L + W), where L
and W are the length and width of the diffraction pattern, respectively. EI values were calculated for shear rates between 0.5 and
15 Pa; an increased EI at a given shear stress indicates greater cell
deformation and hence greater RBC deformability. All measurements were
carried out at 37°C.
Determination of Cytosolic Calcium Concentration
Cytosolic calcium concentration was determined in a separate series of experiments for RBC incubated with L-NAME (103 M) or SNP (106 and 105
M) using a method modified from David-Dufilho et al. (11). RBCs were separated using a density gradient (Histopaque-1077) to
eliminate white blood cell contaminants. The cells were washed twice in
PBS and once in HEPES buffer [containing (in mM) 123 NaCl, 5 KCl, 1 MgCl2 · 6H2O, 1.3 CaCl2, 10 glucose, and 25 HEPES; pH = 7.4] and then resuspended in the
HEPES buffer at a hematocrit of 40%. After the addition of the NO
donor SNP or the nonspecific NOS inhibitor L-NAME, the
suspension hematocrit was immediately reduced to 1%, and RBCs were
incubated at room temperature (22 ± 1°C) for 30 min. Fura 2-AM
at a concentration of 10 nM was added to this suspension and incubated
at 37°C for 30 min (i.e., total incubation time of 60 min). The
suspension was then centrifuged at 350 g for 5 min, and the
packed RBCs were resuspended in PBS to a hemoglobin concentration of
0.05 g/dl. The fluorescence spectrum was measured using an excitation
range of 335-385 nm with emission monitored at 510 nm using a
spectrofluorophotometer (Shimadzu RF-5000; Tokyo, Japan); the ratio of
fluorescence intensities for the fura 2-Ca2+ complex and
the unchelated fura 2 reflects cytosolic calcium concentration.
Determination of RBC Volume and Shape
Mean RBC volume and volume distribution were determined by an orifice-impedance technique using an automated analyzer (ElZone 280 PC, Micromeritics; Norcross, GA). RBCs were also fixed in isotonic 0.5% glutaraldehyde in PBS and kept at room temperature until their shape was examined via phase-contrast or differential interference contrast light microscopy.Statistics
Results are expressed as means ± SE. Statistical comparisons between groups were done by repeated-measures ANOVA, followed by Newman-Keuls posttest, with P values <0.05 accepted as statistically significant.| |
RESULTS |
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TOP
ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
REFERENCES
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RBC Deformability at Various Shear Stresses
RBC deformability (i.e., the EI) for control RBCs was measured at various shear stresses and, as expected (18), was found to increase with increasing applied stress levels (Fig. 1). Figure 1 also presents EI values at various shear stresses for RBCs incubated with L-NAME (103 M), where it can be seen that
L-NAME-treated cells are significantly less deformable at
stress levels of 5 Pa or less: the difference in the EI was more
prominent at the lower stress levels and disappeared at shear stresses
higher than 5 Pa. This pattern of altered RBC deformability (i.e.,
notable effects at lower stress levels) was consistent for all
experimental protocols, and therefore EI values measured at 1.58 Pa
are presented throughout as a measure of RBC deformability.
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Effects of NOS Inhibitors on RBC Deformability
Both the nonspecific NOS inhibitor L-NAME and the specific iNOS inhibitor SMT affected RBC EI in a biphasic manner (Fig. 2). No significant decreases of the EI were observed at concentrations below 103 M for
L-NAME and 104 M for SMT, whereas at these
levels RBC deformability was significantly impaired. Increasing the
L-NAME concentration above 103 M and the SMT
concentration above 104 M did not result in a further
decrease of EI for either agent, but rather the absence of significant
differences and a trend toward control EI levels. Therefore, fixed
concentrations of L-NAME (103 M) and SMT
(104 M) were chosen and used throughout the study.
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Effects of NO Donors on RBC Deformability
RBC deformability was significantly enhanced by SNP at 106 M, with EI values not different from control values
at lower or higher concentrations (Fig.
3A). DETA-NONOate yielded a
similar trend (i.e., maximal effect at 106 M), although
the increase of deformability did not reach significance (Fig.
3B). Note that at higher concentrations the effects of NO donors were not present and RBC deformability tended to decrease.
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Reversal of NOS Inhibitor Effects by NO Donors
The impairment of RBC deformability by 103 M
L-NAME shown in Figs. 1 and 2 was also confirmed in a
separate series of experiments designed to determine the combined
effects of an NOS inhibitor plus an NO donor. As shown in Fig.
4, both NO donors (SNP and DETA-NONOate)
reversed the reduced deformability caused by 103 M
L-NAME in a dose-dependent manner, with the maximum
effective dose greater for SNP (i.e., 105 M for SNP vs.
106 M for DETA-NONOate). Note that even in the presence
of 103 M L-NAME, RBC deformability for cells
incubated with 106 M DETA-NONOate was significantly
enhanced compared with control (Fig. 4B).
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The decreased RBC deformability due to the specific iNOS inhibitor SMT
(104 M) was also reversed by the NO donors SNP and
DETA-NONOate (Fig. 5). SNP at
106 and 105 M significantly reversed the
decrease due to SMT; DETA-NONOate at 106 M yielded the
same result.
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Effects of Soluble Guanylate Cyclase Inhibitors on RBC Deformability
ODQ concentrations of 104-106 M
were employed to determine the effects of this soluble guanylate
cyclase inhibitor on RBC deformability: RBC deformability was reduced
to the same extent at all three concentrations (data not shown), and
thus 105 M was used for subsequent studies. DMSO at a
final concentration identical to that employed as the solvent for ODQ
did not significantly affect RBC deformability.
The decrease of RBC deformability due to 105 M ODQ
was reversed, in a dose-dependent manner, by the NO donors SNP and
DETA-NONOate (Fig. 6), with SNP requiring
a higher level for maximal EI. This series of experiments was repeated
using a higher concentration of ODQ (104 M), with the
results indicating a pattern similar to the effects of SNP and
DETA-NONOate shown in Fig. 6. Thus the reversal of the ODQ effects at
both 105 and 104 M ODQ essentially
eliminates the possibility that this reversal pattern results from an
inadequate inhibition of guanylate cyclase.
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In a separate set of experiments (n = 5)
conducted using 105 M methylene blue, an inhibitor
of both NOS and soluble guanylate cyclase, a significant impairment in
the RBC EI was observed. This decrease of the EI was prevented by SNP,
with a dose-dependent pattern similar to that observed with ODQ (data
not shown).
Effects of the NO Precursor L-Arginine
The decreases of RBC deformability due to the NOS inhibitors L-NAME (103 M) and SMT (104 M)
and the soluble guanylate cyclase inhibitor ODQ (105 M)
were prevented by the NO precursor L-arginine at
105 M (Fig. 7). As shown in
Fig. 7, L-arginine alone had no effect on RBC
deformability.
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Role of Potassium Channel Blockade
In a preliminary series of experiments, the nonspecific potassium channel blocker TEA was found to modestly enhance RBC deformability at concentrations between 104 and 107 M,
presumably due to slight cell swelling and the consequent reduction of
cytoplasmic viscosity (data not shown). The highest concentration of
TEA that did not affect the RBC EI after 1-h incubation was found to be
108 M. Notably, however, TEA at this concentration
prevented the impairment of RBC deformability induced by
103 M L-NAME (Fig.
8).
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Intracellular Calcium Concentration, RBC Shape, and RBC Volume
Neither SNP (106 or 105 M) nor
L-NAME (103 M) affected intracellular calcium
concentration (P > 0.1). At the concentrations used herein, neither RBC volume nor shape were affected by the NO donors, NOS inhibitors, or soluble guanylate cyclase inhibitors.
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DISCUSSION |
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TOP
ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
REFERENCES
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The ability of the entire RBC to deform (i.e., RBC deformability) is of crucial importance for the maintenance of normal circulation: it allows the passage of erythrocytes through narrow capillaries in the microcirculation and reduces blood viscosity at high shear rates in large blood vessels (10, 40). With the use of a rat cremaster muscle preparation, Lipowsky and co-workers (26, 35) demonstrated that decreased RBC deformability results in impaired microcirculatory perfusion. The major determinants of RBC deformability are cell geometry (i.e., membrane surface area-to-volume ratio), cell shape, cytoplasmic viscosity, and membrane mechanical properties. Although both the lipid bilayer and the RBC cytoskeleton may affect RBC membrane mechanical behavior, the physical properties of the membrane skeleton are the main determinants of the viscous and elastic characteristics of the membrane (30, 31, 40). Maintenance of normal RBC deformability depends critically on the metabolic state of the cell: 1) metabolic energy is necessary for the proper operation of ion pumps (e.g., Na+-K+-ATPase and Ca2+-ATPase); 2) Na+-K+-ATPase is the primary regulator of RBC volume and hence cytoplasmic viscosity via maintaining the osmotic balance across the cell membrane (32); and 3) Ca2+-ATPase maintains the low intracellular Ca2+ concentration essential for normal RBC deformability (33, 34).
The results of the current study confirm and extend prior reports (3, 6, 21, 22, 29, 41, 42) indicating that NO plays a role in maintaining RBC deformability. NO, at some critical concentration, therefore appears necessary for preserving this cellular mechanical property, and thus its role can be postulated as "regulatory" vis-a-vis normal RBC deformability. In support of this suggestion are our observations that inhibition of NOS, by both nonspecific (L-NAME; Figs. 1 and 2) and iNOS-specific (SMT; Fig. 2) inhibitors, resulted in significant impairment of RBC deformability, which could be restored by external NO donors (SNP and DETA-NONOate; Figs. 4 and 5).
The effects of both specific iNOS and nonspecific NOS inhibitors could also be reversed by the NOS substrate L-arginine (Fig. 7), indicating the role of internally synthesized NO in the regulation of RBC mechanical properties. RBCs are exposed to NO synthesized by other cells in their natural environment, such as endothelial cells and granulocytes (5, 27), but also contain systems to synthesize NO internally (9, 17). Jubelin et al. (17) demonstrated that RBCs are positive for iNOS (NOS2) and eNOS (NOS2), whereas Chen and Mehta (9) reported that erythrocytes exhibit eNOS but not iNOS activity. Kang et al. (19) indicated that human RBCs collected from normal adults possess both inducible and endothelial isoforms of NOS but that these proteins are without catalytic activity. In contrast with Kang et al.'s report (19), the results of this study indicated that iNOS may play role in maintaining normal RBC deformability.
In the present study, impaired RBC deformability due to L-NAME was restored by two NO donors (i.e., SNP and DETA-NONOate) having different chemical structures, thus minimizing the possibility of an effect unrelated to NO release. Additional support for a specific NO effect emerges from the results shown in Figs. 4-6, where it can be seen that the optimal concentration of SNP was higher than that for DETA-NONOate. This difference of optimal concentration most likely relates to the release of two NO molecules for each DETA-NONOate (20) versus only one for SNP, thereby yielding twice the NO level at a given molar concentration. It is interesting to note that NO has biphasic effects on RBC mechanical properties, with this effect especially obvious in the experiments where endogenous NO synthesis was inhibited (Figs. 4 and 5). RBC deformability was found to be maximal at a given concentration that depended on the NO-releasing capacity of the specific NO donor; above this concentration red cell deformability was impaired. The biphasic nature of this effect again suggests that the presence of NO at a critical concentration is crucial for the maintenance of normal RBC deformability and that at higher concentrations NO adversely affects cellular mechanical behavior (6, 21). The reason for the biphasic effect is not yet certain but may relate to the known pathophysiological effects induced by higher concentrations of NO or its metabolites, such as increased oxidant stress (14, 28, 44).
Although both our findings and literature reports (e.g., 3, 6, 21, 22, 29, 41, 42) demonstrate the role of NO in regulating RBC mechanical
properties, the mechanism(s) responsible for the effects of NO on RBC
deformability has yet to be fully defined. Classically, the effects of
NO are known to be mediated by cGMP produced by soluble guanylate
cyclase, and Petrov et al. (37, 38) demonstrated the
existence of soluble guanylate cyclase in RBCs. In support of a role
for guanylate cyclase in the regulatory pathway of RBC deformability by
NO, it is notable that soluble guanylate cyclase inhibitors (i.e., ODQ
and methylene blue) impaired RBC deformability (Fig. 6). However, this
effect could be reserved by the NO donors SNP or DETA-NONOate in a
dose-dependent manner (Fig. 6). If cGMP was the only mediator of the NO
effect, the decrease of RBC deformability by guanylate cyclase
inhibitors would not have been reversed by these NO donors
(13). Furthermore, the dose-dependent reversal of RBC
deformability with these NO donors in the presence of 105
M ODQ was also evident at a higher concentration (104 M),
thereby essentially excluding the possibility of only partial inhibition of guanylate cyclase.
These above-mentioned findings suggest that the regulatory effect of NO on erythrocyte deformability is only partially mediated by soluble guanylate cyclase and that other mechanisms need to be considered. For example, there are several reports indicating the cGMP-independent effects of NO in various tissues. Ahmmed et al. (2) demonstrated that NO modulates Na+ current in guinea pig and mouse ventricular myocytes via both cGMP- and cAMP-dependent pathways (2), and Pinilla et al. (39) suggested that NO stimulates growth hormone secretion through a specific calcium-cGMP-independent mechanism. It is known that NO has two different effects on megakaryocytes: one of them, the induction of platelet formation, is cGMP independent, whereas the second one, the stimulation of apoptosis, is cGMP dependent (4). Other signal transduction pathways may also be involved, with Oonishi et al. (34) suggesting an important role for cAMP in the maintenance of normal RBC deformability.
Ion transport across the RBC membrane is influenced by NO
(36-38), and it has been demonstrated that both the
Na+-K+-ATPase and Ca2+-ATPase
activities of RBCs are stimulated by NO donors (12, 36).
However, in the present study, intracellular calcium concentration was
not altered by either the NOS inhibitor L-NAME or the NO
donor SNP. NO is also known to affect potassium transport across the membrane, either directly or indirectly (e.g., mediated by nitrite or
peroxynitrite) (1, 8, 15). Although potassium currents were not measured in this study, it was observed that blockade of
potassium transport through the RBC membrane prevented the adverse
effect of L-NAME on RBC deformability (Fig. 8). This
finding suggests that the inhibition of NO synthesis by the nonspecific NOS inhibitor L-NAME may lead to deterioration of RBC
mechanical properties by increasing the potassium permeability of the
RBC membrane. L-Arginine competitive analogs have been
shown to increase RBC potassium efflux and to decrease intracellular NO
generation (8), thus supporting this suggestion. However,
increased potassium release from RBCs also occurs under the influence
of NO metabolites (e.g., nitrite and peroxynitrite) (1,
15). Altered potassium transport might be expected to influence
RBC shape and volume due to osmotic gradients; however, in our study,
neither RBC shape nor volume were altered. Our morphological results
are thus consistent with those of Walter et al. (43), who
indicated that SNP induced reversible echinocytic shape transformation
only at very high levels (102 M), whereas at lower
concentrations SNP failed to alter RBC shape.
In summary, the results of this study clearly indicate that NO can markedly affect RBC deformability and therefore suggest the regulatory role of NO in maintaining normal RBC deformability. Under in vivo conditions, RBC are exposed to two sources of NO: one synthesized by endothelial cells, which diffuses into the bloodstream, and one synthesized within the RBC itself. Thus, in addition to the well-known effect of NO on vascular hindrance, the effect of NO on RBC deformability appears to have a direct role in the control of blood flow. Unfortunately, the target structure or function for the effects of NO on RBC mechanical behavior have not yet been established; although the most probable site for this effect is altered phosphorylation of the RBC cytoskeleton (34), further studies are required to clarify this aspect.
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ACKNOWLEDGEMENTS |
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This study was carried out as part of a PhD thesis by M. Bor-Kucukatay presented to Akdeniz University Health Sciences Institute.
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FOOTNOTES |
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This study was supported by a grant from North Atlantic Treaty Organization Science Fellowships Program by The Scientific and Technical Research Council of Turkey, by National Heart, Lung, and Blood Institute Research Grants HL-15722 and HL-48484, Fogarty International Research Collaboration Award IR03-TWO1295, and by Akdeniz University Research Fund 20.01.0122.03.
Address for reprint requests and other correspondence: O. K. Baskurt, Dept. of Physiology, Akdeniz Univ. School of Medicine, Antalya, 07070 Turkey (E-mail: baskurt{at}akdeniz.edu.tr).
The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
First published January 9, 2003;10.1152/ajpheart.00665.2002
Received 30 July 2002; accepted in final form 2 January 2003.
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REFERENCES |
|---|
|
TOP
ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
REFERENCES
|
|---|
1.
Adragna, NC,
and
Lauf PK.
Role of nitrite, a nitric oxide derivative, in K-Cl cotransport activation of low-potassium sheep red blood cells.
J Membr Biol
166:
157-167,
1998[Web of Science][Medline].
2.
Ahmmed, GU,
Xu Y,
Hong Dong P,
Zhang Z,
Eiserich J,
and
Chiamvimonvat N.
Nitric oxide modulates cardiac Na+ channel via protein kinase A and protein kinase G.
Circ Res
89:
1005-1013,
2001
3.
Bateman, RM,
Jagger JE,
Sharpe MD,
Ellsworth ML,
Mehta S,
and
Ellis CG.
Erythrocyte deformability is a nitric oxide-mediated factor in decreased capillary density during sepsis.
Am J Physiol Heart Circ Physiol
280:
H2848-H2856,
2001
4.
Battinelli, E,
Willoughby SR,
Foxall T,
Valeri CR,
and
Loscalzo J.
Induction of paltelet formation from megakaryocytoid cells by nitric oxide.
Proc Natl Acad Sci USA
98:
14458-14463,
2001
5.
Bhagat, K,
and
Vallance P.
Nitric oxide 9 years on.
J R Soc Med
89:
667-673,
1996[Web of Science][Medline].
6.
Bor-Kucukatay, M,
Yalcin O,
Gokalp O,
Kipmen-Korgun D,
Yesilkaya A,
Baykal A,
Ispir M,
Senturk UK,
Kaputlu I,
and
Baskurt OK.
Red blood cell rheological alterations in hypertension induced by chronic inhibition of nitric oxide synthesis in rats.
Clin Hemorheol
22:
267-275,
2000.
7.
Butler, AR,
Megson IL,
and
Wright PG.
Diffusion of nitric oxide and scavenging by blood in the vasculature.
Biochim Biophys Acta
1425:
168-176,
1998[Medline].
8.
Caramelo, C,
Riesco A,
Outeirino J,
Milas I,
Blum G,
Monzu B,
Casado V,
Sanchez L,
Mosquera JR,
Casado S,
and
Farre AL.
Effects of nitric oxide on red blood cell cell: changes in erythrocyte resistance to hypotonic hemolysis and potassium efflux by experimental maneuvers that decrease nitric oxide.
Biochem Biophys Res Commun
15:
447-454,
1994.
9.
Chen, LY,
and
Mehta JL.
Evidence for the presence of L-arginine-nitric oxide pathway in human red blood cells: relevance in the effects of red blood cells on platelet function.
J Cardiovasc Pharmacol
32:
57-61,
1998[Web of Science][Medline].
10.
Chien, S.
Red cell deformability and its relevance to blood flow.
Annu Rev Physiol
49:
177-192,
1987[Web of Science][Medline].
11.
David-Dufilho, M,
Montenay-Grestier T,
and
Devynck MA.
Fluorescence measurements of free Ca2+ concentration in human erythrocytes using the Ca2+-indicator fura-2.
Cell Calcium
9:
167-179,
1988[Web of Science][Medline].
12.
De Oliveira Elais, M,
Tavares de Lima W,
Vannuchi YB,
Marcourakis T,
da Silva ZL,
Trezena AG,
and
Scavone C.
Nitric oxide modulates Na+, K+-ATPase activity through cyclic GMP pathway in proximal rat trachea.
Eur J Pharmacol
367:
307-314,
1999[Web of Science][Medline].
13.
Denninger, JW,
and
Marletta MA.
Guanylate cyclase and the NO/cGMP signaling pathway.
Biochim Biophys Acta
1411:
334-350,
1999[Medline].
14.
Espey, MG,
Miranda KM,
Thomas DD,
Xavier S,
Citrin D,
Vitek MP,
and
Wink DA.
A chemical perspective on the interplay between NO, reactive oxygen species and reactive nitrogen species.
Ann NY Acad Sci
962:
194-206,
2002.
15.
Grzelak, A,
Mazur J,
and
Bartosz G.
Peroxynitrate activates K+-Cl cotransport in human erythrocytes.
Cell Biol Int
25:
1163-1165,
2001[Web of Science][Medline].
16.
Jia, L,
Bonaventura C,
Bonaventura J,
and
Stamler JS.
S-nitrosohaemoglobin: a dynamic activity of blood involved in vascular control.
Nature
380:
221-226,
1996[Medline].
17.
Jubelin, BC,
and
Gierman JL.
Erythrocytes may synthesize their own nitric oxide.
Am J Hypertens
9:
1214-1219,
1996[Web of Science][Medline].
18.
Hardeman, MR,
Goedhart PT,
Dobbe JGG,
and
Lettinga KP.
Laser assisted optical rotational cell analyzer (LORCA): a new instrument for measurement of various structural hemorhelogical parameters.
Clin Hemorheol
14:
605-618,
1994[Web of Science].
19.
Kang, ES,
Ford K,
Grokulsky G,
Wang YB,
Chiang TM,
and
Acchiardo SR.
Normal circulating adult human red blood cells contain inactive NOS proteins.
J Lab Clin Med
135:
444-451,
2000[Web of Science][Medline].
20.
Keefer, LK,
Nims RW,
Davies KM,
and
Wink DA.
"NONOates" (1-substituted diazen-1-ium-1,2-diolates) as nitric oxide donors: convenient nitric oxide dosage forms.
Methods Enzymol
268:
281-293,
1996[Web of Science][Medline].
21.
Korbut, R,
and
Gryglewski RJ.
Nitric oxide from polymorphonuclear leukocytes modulates red blood cell deformability in vitro.
Eur J Pharmacol
234:
17-22,
1993[Web of Science][Medline].
22.
Korbut, R,
and
Gryglewski RJ.
The effect of prostacyclin and nitric oxide on deformability of red blood cells in septic shock in rats.
J Physiol Pharmacol
47:
591-599,
1996[Web of Science][Medline].
23.
Kon, K,
Maeda N,
and
Shiga T.
Effect of nitric oxide on the oxygen transport of human erythrocytes.
J Toxicol Environ Health
2:
1109-1113,
1997.
24.
Liao, JC,
Hein TW,
Vaughn MW,
Huang KT,
and
Kuo L.
Intravascular flow decreases erythrocyte consumption of nitric oxide.
Proc Natl Acad Sci USA
96:
8757-8761,
1999
25.
Lloyd-Jones, DM,
and
Bloch KD.
The vascular biology of nitric oxide and its role in atherogenesis.
Annu Rev Med
47:
365-375,
1996[Web of Science][Medline].
26.
Lipowsky, HH,
Cram LE,
Justice W,
and
Eppihimer MJ.
Effect of erythrocyte deformability on in vivo red cell transit time and hematocrit and their correlation with in vitro filterability.
Microvasc Res
46:
43-64,
1993[Web of Science][Medline].
27.
Marin, J,
and
Rodriguez-Martinez MA.
Role of vascular nitric oxide in physiological and pathological conditions.
Pharmacol Ther
75:
111-134,
1997[Web of Science][Medline].
28.
May, ZM,
Qu ZC,
Xia L,
and
Cobb CE.
Nitrite uptake and metabolism and oxidant stress in human erythrocytes.
Am J Physiol Cell Physiol
279:
C1946-C1954,
2000
29.
Mesquita, R,
Pires I,
Saldanha C,
and
Martins-Silva J.
Effects of acetylcholine and spermineNONOate on erythrocyte hemorheologic and oxygen carrying properties.
Clin Hemorheol
25:
153-163,
2001.
30.
Mohandas, N,
and
Chasis JA.
Red blood cell deformability, membrane material properties and shape: regulation by transmembrane, skeletal and cytosolic proteins and lipids.
Semin Hematol
30:
171-192,
1983.
31.
Mohandas, N,
Chasis JA,
and
Shohet SB.
The influence of membrane skeleton on red cell deformability, membrane material properties, and shape.
Semin Hematol
20:
225-242,
1983[Web of Science][Medline].
32.
Mohandas, N,
and
Shohet SB.
The role of membrane-associated enzymes in regulation of erythrocyte shape and deformability.
Clin Haematol
10:
223-237,
1981[Web of Science][Medline].
33.
Murakami, J,
Maeda N,
Kon K,
and
Shiga T.
A contribution of calmodulin to cellular deformability of calcium-loaded human erythrocytes.
Biochim Biophys Acta
863:
23-32,
1986[Medline].
34.
Oonishi, T,
Sakashita K,
and
Uyesaka N.
Regulation of red blood cell filterability by Ca2+ influx and cAMP-mediated signaling pathways.
Am J Physiol Cell Physiol
273:
C1828-C1834,
1997
35.
Pathasarathi, K,
and
Lipowsky HH.
Capillary recruitment in response to tissue hypoxia and its dependence on red blood cell deformability.
Am J Physiol Heart Circ Physiol
277:
H2145-H2157,
1999
36.
Pernollet, MG,
Lantoine F,
and
Devynck MA.
Nitric oxide inhibits ATP-dependent Ca2+ uptake into platelet membrane vesicles.
Biochem Biophys Res Commun
222:
780-785,
1996[Web of Science][Medline].
37.
Petrov, V,
Amery A,
and
Lijnen P.
Role of cyclic GMP in atrial-natriuretic-peptide stimulation of erythrocyte Na+/H+ exchange.
Eur J Biochem
221:
195-199,
1994[Web of Science][Medline].
38.
Petrov, V,
and
Lijnen P.
Regulation of human erythrocyte Na+/H+ exchange by soluble and particulate guanylate cyclase.
Am J Physiol Cell Physiol
271:
C1556-C1564,
1996
39.
Pinilla, L,
Tena-Sempere M,
and
Aguilar E.
Nitric oxide stimulates growth hormone secretion in vitro through a calciumand cyclic guanosine monophosphate-independent mechanism.
Horm Res
51:
242-247,
1999[Web of Science][Medline].
40.
Shiga, T,
Maeda N,
and
Kon K.
Erythrocyte rheology.
Crit Rev Oncol Hematol
10:
9-48,
1990[Web of Science][Medline].
41.
Starzyk, D,
Korbut R,
and
Gryglewski RJ.
The role of nitric oxide in regulation of deformability of red blood cells in acute phase of endotoxaemia in rats.
J Physiol Pharmacol
48:
731-735,
1997[Web of Science][Medline].
42.
Starzyk, D,
Korbut R,
and
Gryglewski RJ.
Effects of nitric oxide and prostacyclin on deformability and aggregability of red blood cells of rats ex vivo and in vitro.
J Physiol Pharmacol
50:
629-637,
1999[Web of Science][Medline].
43.
Walter, R,
Mark M,
Gaudenz R,
Harris LG,
and
Reinhart WH.
Influence of nitrosovasodilators and endothelin-1 on rheology of human blood in vitro.
Br J Pharmacol
128:
744-750,
1999[Web of Science][Medline].
44.
Wink, DA,
Miranda KM,
and
Espsy MG.
Cytotoxicity related to oxidative and nitrosative stress by nitric oxide.
Exp Biol Med (Maywood)
226:
621-623,
2001
45.
Yonetani, T.
Nitric oxide and hemoglobin.
Nippon Yakurigaku Zasshi
112:
155-160,
1998[Medline].
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P < 0.01, and
fP < 0.001, significantly
different from control.
