Abstract
Renal ischemia rapidly mobilizes endothelial progenitor cells (EPCs), which provides renoprotection in acute kidney injury (AKI). Indoxyl sulfate (IS) is a protein-binding uremic toxin with a potential role in endothelial injury. In this study, we examined the effects and mechanisms of action of IS on EPCs in AKI. Forty-one consecutive patients (26 male; age, 70.1 ± 14.1 years) diagnosed with AKI according to the AKIN criteria were enrolled. The AKI patients had higher serum IS levels than patients with normal kidney function (1.35 ± 0.94 × 10−4M vs. 0.02 ± 0.02 × 10−4M, P < 0.01). IS levels were negatively correlated to the number of double-labeled (CD34+KDR+) circulating EPCs (P < 0.001). After IS stimulation, the cells displayed decreased expression of phosphorylated endothelial nitric oxide (NO) synthase, vascular cell adhesion molecule-1, increased reactive oxygen species, decreased proliferative capacity, increased senescence and autophagy, as well as decreased migration and angiogenesis. These effects of IS on EPCs were reversed by atorvastatin. Further, exogenous administration of IS significantly reduced EPC number in Tie2-GFP transgenic mice and attenuated NO signaling in aortic and kidney arteriolar endothelium after kidney ischemia–reperfusion injury in mice, and these effects were restored by atorvastatin. Our results are the first to demonstrate that circulating IS is elevated in AKI and has direct effects on EPCs via NO-dependent mechanisms both in vitro and in vivo. Targeting the IS-mediated pathways by NO-releasing statins such as atorvastatin may preempt disordered vascular wall pathology, and represent a novel EPC-rescued approach to impaired neovascularization after AKI.
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Abbreviations
- AKI:
-
Acute kidney injury
- APO:
-
Apocynin
- ATO:
-
Atorvastatin
- CKD:
-
Chronic kidney disease
- CASP3:
-
Activated caspase 3
- Ctrl:
-
Control
- DAPI:
-
DNA fluorochrome 4′-6-diamidine-2-phenyl indole
- DCFDA:
-
Dichlorofluorescin diacetate
- eNOs:
-
Endothelial nitric oxide synthase
- EPC:
-
Endothelial progenitor cells
- ESRD:
-
End stage renal disease
- FITC:
-
Fluorescein isothiocyanate
- GAM:
-
Generalized additive model
- ICAM-1:
-
Intercellular adhesion molecule 1
- IS:
-
Indoxyl sulfate
- LC3:
-
Light Chain 3
- MTT:
-
3-(4,5-cimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide
- NAC:
-
N-acetylcsyteine
- NO:
-
Nitric oxide
- NOx:
-
Reactive nitrogen species
- NSARF:
-
National Taiwan University Hospital Study Group on Acute Renal Failure
- OAT:
-
Organic anion transporter
- ROS:
-
Reactive oxygen species
- SNP:
-
Sodium nitroprusside
- Prob:
-
Probenecid
- TNF:
-
Transforming growth factor
- TUNEL:
-
Terminal deoxyribonucleotidyl transferase (TDT)-mediated dUTP-digoxigenin nick end labeling
- VCAM:
-
Vascular cell adhesion molecule
- VWF:
-
Von Willebrand factor
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Acknowledgments
The authors would like to thank the staff of the Second Core Lab of the Department of Medical Research in the National Taiwan University Hospital for technical assistance. This study was supported by The Ta-Tung Kidney Foundation, Taiwan National Science Council (Grant NSC 101-2314-B-002-132-MY3, Grant NSC 101-2314-B-002-085-MY3, and Grant NSC 100-2314-B-002-119-), 100-N1776 from NTUH, 101-M1953, 102-S2097 and NTUH-TVGH Joint Research Program (VN9803, VN9906 and VN10009).
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Vin-Cent Wu and Guang-Huar Young contributed equally to this work.
ClinicalTrials#. Gov number, NCT00451373.
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Wu, VC., Young, GH., Huang, PH. et al. In acute kidney injury, indoxyl sulfate impairs human endothelial progenitor cells: modulation by statin. Angiogenesis 16, 609–624 (2013). https://doi.org/10.1007/s10456-013-9339-8
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DOI: https://doi.org/10.1007/s10456-013-9339-8