title: Indolic uremic solutes enhance procoagulant activity of red blood cells through phosphatidylserine exposure and microparticle release.
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.
authors.affiliation: Department of Medical Laboratory Science and Technology, Harbin Medical University-Daqing, 39 Xinyang Road, Gaoxin District, Daqing 163319, China. gaochunyan1234@163.com.
authors.forename: Chunyan
authors.initials: C
authors.lastname: Gao
authors.affiliation: Department of Medical Laboratory Science and Technology, Harbin Medical University-Daqing, 39 Xinyang Road, Gaoxin District, Daqing 163319, China. jishuting@tom.com.
authors.forename: Shuting
authors.initials: S
authors.lastname: Ji
authors.affiliation: Department of General Surgery, The Fifth Hospital. dongweijun19771977@163.com.
authors.forename: Weijun
authors.initials: W
authors.lastname: Dong
authors.affiliation: Department of Medical Laboratory Science and Technology, Harbin Medical University-Daqing, 39 Xinyang Road, Gaoxin District, Daqing 163319, China. jianyanzhongxin@126.com.
authors.forename: Yushan
authors.initials: Y
authors.lastname: Qi
authors.affiliation: Department of Medical Laboratory Science and Technology, Harbin Medical University-Daqing, 39 Xinyang Road, Gaoxin District, Daqing 163319, China. fyzqx@sina.com.
authors.forename: Wen
authors.initials: W
authors.lastname: Song
authors.affiliation: Department of Medical Laboratory Science and Technology, Harbin Medical University-Daqing, 39 Xinyang Road, Gaoxin District, Daqing 163319, China. cdb1024@163.com.
authors.forename: Debin
authors.initials: D
authors.lastname: Cui
authors.affiliation: Department of Hematology, The First Hospital, Harbin Medical University, 23 Youzheng Street, Nangang District, Harbin 150001, China. jialan_shi@hms.harvard.edu.|Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02132, USA. jialan_shi@hms.harvard.edu.
authors.forename: Jialan
authors.initials: J
authors.lastname: Shi
mesh_terms: D000925:Anticoagulants / Q000494:pharmacology; D001777:Blood Coagulation / Q000187:drug effects*; D002118:Calcium / Q000097:blood; D055252:Cell-Derived Microparticles / Q000378:metabolism*; D003600:Cytosol / Q000378:metabolism; D004912:Erythrocytes / Q000187:drug effects*; D015951:Factor Xa / Q000032:analysis; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D007200:Indican / Q000494:pharmacology / Q000652:urine; D007210:Indoleacetic Acids / Q000494:pharmacology / Q000652:urine; D007211:Indoles / Q000494:pharmacology* / Q000652:urine; D008297:Male; D010718:Phosphatidylserines / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D013925:Thromboplastin / Q000032:analysis; D013927:Thrombosis / Q000097:blood / Q000652:urine; D014511:Uremia / Q000378:metabolism*; D055815:Young Adult
chemical_list: D000925:Anticoagulants; D007210:Indoleacetic Acids; D007211:Indoles; D010718:Phosphatidylserines; C030737:indoleacetic acid; D013925:Thromboplastin; D015951:Factor Xa; D007200:Indican; D002118:Calcium
keywords: indole-3-acetic acid; indoxyl sulfate; microparticle; phosphatidylserine; procoagulant; red blood cell