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Indolic uremic solutes enhance procoagulant activity of red blood cells through phosphatidylserine exposure and microparticle release.

Gao C, Ji S, Dong W, Qi Y, Song W, Cui D, Shi J - Toxins (Basel) (2015)

Bottom Line: However, the effect of uremic solutes indoxyl sulfate (IS) and indole-3-acetic acid (IAA) on procoagulant activity (PCA) of erythrocyte is unclear.Our results suggest that RBC eryptosis in uremic solutes IS and IAA plays an important role in thrombus formation through releasing RMPs and exposing PS.Lactadherin acts as an efficient anticoagulant in this process.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Laboratory Science and Technology, Harbin Medical University-Daqing, 39 Xinyang Road, Gaoxin District, Daqing 163319, China. gaochunyan1234@163.com.

ABSTRACT
Increased accumulation of indolic uremic solutes in the blood of uremic patients contributes to the risk of thrombotic events. Red blood cells (RBCs), the most abundant blood cells in circulation, may be a privileged target of these solutes. However, the effect of uremic solutes indoxyl sulfate (IS) and indole-3-acetic acid (IAA) on procoagulant activity (PCA) of erythrocyte is unclear. Here, RBCs from healthy adults were treated with IS and IAA (mean and maximal concentrations reported in uremic patients). Phosphatidylserine (PS) exposure of RBCs and their microparticles (MPs) release were labeled with Alexa Fluor 488-lactadherin and detected by flow cytometer. Cytosolic Ca(2+) ([Ca(2+)]) with Fluo 3/AM was analyzed by flow cytometer. PCA was assessed by clotting time and purified coagulation complex assays. We found that PS exposure, MPs generation, and consequent PCA of RBCs at mean concentrations of IS and IAA enhanced and peaked in maximal uremic concentrations. Moreover, 128 nM lactadherin, a PS inhibitor, inhibited over 90% PCA of RBCs and RMPs. Eryptosis or damage, by indolic uremic solutes was due to, at least partially, the increase of cytosolic [Ca(2+)]. Our results suggest that RBC eryptosis in uremic solutes IS and IAA plays an important role in thrombus formation through releasing RMPs and exposing PS. Lactadherin acts as an efficient anticoagulant in this process.

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Effect of Ca2+ withdrawal on IS and IAA induced PS exposure. The percentage of lactadherin binding erythrocytes after a 24 h treatment with IS (0.1 mM) or IAA (20 μM) in the presence and absence of EGTA (1 mM). KCl or ethanol was utilized as their respective controls. Data are displayed as mean ± SD for triplicate samples of independent experiments, * indicate p < 0.001.
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toxins-07-04390-f003: Effect of Ca2+ withdrawal on IS and IAA induced PS exposure. The percentage of lactadherin binding erythrocytes after a 24 h treatment with IS (0.1 mM) or IAA (20 μM) in the presence and absence of EGTA (1 mM). KCl or ethanol was utilized as their respective controls. Data are displayed as mean ± SD for triplicate samples of independent experiments, * indicate p < 0.001.

Mentions: In order to verify whether the PS exposure on erythrocyte by indolic uremic solutes was secondary to an increase of [Ca2+], erythrocytes were exposed to IS (0.1 mM) and IAA (20 μM) for 24 h either in the presence of extracellular Ca2+ (1 mM) or in the nominal absence of Ca2+ and presence of the Ca2+ chelator EGTA (1 mM). As shown in Figure 3, the effect of IS and IAA on the increased PS exposure was virtually abolished in the absence of Ca2+.


Indolic uremic solutes enhance procoagulant activity of red blood cells through phosphatidylserine exposure and microparticle release.

Gao C, Ji S, Dong W, Qi Y, Song W, Cui D, Shi J - Toxins (Basel) (2015)

Effect of Ca2+ withdrawal on IS and IAA induced PS exposure. The percentage of lactadherin binding erythrocytes after a 24 h treatment with IS (0.1 mM) or IAA (20 μM) in the presence and absence of EGTA (1 mM). KCl or ethanol was utilized as their respective controls. Data are displayed as mean ± SD for triplicate samples of independent experiments, * indicate p < 0.001.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4663509&req=5

toxins-07-04390-f003: Effect of Ca2+ withdrawal on IS and IAA induced PS exposure. The percentage of lactadherin binding erythrocytes after a 24 h treatment with IS (0.1 mM) or IAA (20 μM) in the presence and absence of EGTA (1 mM). KCl or ethanol was utilized as their respective controls. Data are displayed as mean ± SD for triplicate samples of independent experiments, * indicate p < 0.001.
Mentions: In order to verify whether the PS exposure on erythrocyte by indolic uremic solutes was secondary to an increase of [Ca2+], erythrocytes were exposed to IS (0.1 mM) and IAA (20 μM) for 24 h either in the presence of extracellular Ca2+ (1 mM) or in the nominal absence of Ca2+ and presence of the Ca2+ chelator EGTA (1 mM). As shown in Figure 3, the effect of IS and IAA on the increased PS exposure was virtually abolished in the absence of Ca2+.

Bottom Line: However, the effect of uremic solutes indoxyl sulfate (IS) and indole-3-acetic acid (IAA) on procoagulant activity (PCA) of erythrocyte is unclear.Our results suggest that RBC eryptosis in uremic solutes IS and IAA plays an important role in thrombus formation through releasing RMPs and exposing PS.Lactadherin acts as an efficient anticoagulant in this process.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Laboratory Science and Technology, Harbin Medical University-Daqing, 39 Xinyang Road, Gaoxin District, Daqing 163319, China. gaochunyan1234@163.com.

ABSTRACT
Increased accumulation of indolic uremic solutes in the blood of uremic patients contributes to the risk of thrombotic events. Red blood cells (RBCs), the most abundant blood cells in circulation, may be a privileged target of these solutes. However, the effect of uremic solutes indoxyl sulfate (IS) and indole-3-acetic acid (IAA) on procoagulant activity (PCA) of erythrocyte is unclear. Here, RBCs from healthy adults were treated with IS and IAA (mean and maximal concentrations reported in uremic patients). Phosphatidylserine (PS) exposure of RBCs and their microparticles (MPs) release were labeled with Alexa Fluor 488-lactadherin and detected by flow cytometer. Cytosolic Ca(2+) ([Ca(2+)]) with Fluo 3/AM was analyzed by flow cytometer. PCA was assessed by clotting time and purified coagulation complex assays. We found that PS exposure, MPs generation, and consequent PCA of RBCs at mean concentrations of IS and IAA enhanced and peaked in maximal uremic concentrations. Moreover, 128 nM lactadherin, a PS inhibitor, inhibited over 90% PCA of RBCs and RMPs. Eryptosis or damage, by indolic uremic solutes was due to, at least partially, the increase of cytosolic [Ca(2+)]. Our results suggest that RBC eryptosis in uremic solutes IS and IAA plays an important role in thrombus formation through releasing RMPs and exposing PS. Lactadherin acts as an efficient anticoagulant in this process.

Show MeSH
Related in: MedlinePlus