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In acute kidney injury, indoxyl sulfate impairs human endothelial progenitor cells: modulation by statin

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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).

Conflict of interest

The authors have nothing to disclose.

Author information

Authors and Affiliations

  1. Department of Internal Medicine, National Taiwan University Hospital, 7 Chung-Shan South Road, Taipei, 100, Taiwan

    Vin-Cent Wu, Jou-Han Chen, Yung-Ming Chen & Kwan-Dun Wu

  2. Department of Surgery, National Taiwan University Hospital, Taipei, Taiwan

    Guang-Huar Young & Wen-Jo Ko

  3. Division of Cardiology, Department of Internal Medicine, Taipei Veterans General Hospital, Taipei, Taiwan

    Po-Hsun Huang & Jaw-Wen Chen

  4. Department of Laboratory Medicine, National Taiwan University Hospital, Taipei, Taiwan

    Shyh-Chyi Lo

  5. Department of Neurology, National Taiwan University Hospital, Taipei, Taiwan

    Kuo-Chuan Wang

  6. Division of Nephrology, Chang Gung Memorial Hospital, Keelung, Taiwan

    Chiao-Yin Sun

  7. Department of Anatomy and Cell Biology, College of Medicine, National Taiwan University Hospital, Taipei, Taiwan

    Chan-Jung Liang & Yuh-Lien Chen

  8. Yun-Lin Branch, National Taiwan University Hospital, Taipei, Taiwan

    Tao-Ming Huang

  9. Chu-Tung Branch, National Taiwan University Hospital, Taipei, Taiwan

    Fan-Chi Chang

  10. Bei-Hu branch, National Taiwan University Hospital, Taipei, Taiwan

    Yih-Shing Kuo

  11. Division of Nephrology, Chang Gung Memorial Hospital, Kaohsiung, Taiwan

    Jin-Bor Chen

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  1. Vin-Cent Wu
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The NSARF group

Corresponding author

Correspondence to Yung-Ming Chen.

Additional information

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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