Mesenchymal Stem Cell-conditioned Medium Protecting Renal Tubular Epithelial Cells by Inhibiting Hypoxia-inducible Factor-1α and Nuclear Receptor Coactivator-1


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Abstract

Background:Acute kidney injury (AKI) is characterized by inflammatory infiltration and damage and death of renal tubular epithelial cells (RTECs), in which hypoxia plays an important role. Deferoxamine (DFO) is a well-accepted chemical hypoxia-mimetic agent. Mesenchymal stem cell-conditioned medium (MSC-CM) can reduce local inflammation and repair tissue. In this study, we explored the effect and molecular mechanism of MSC-CM-mediated protection of RTECs under DFO-induced hypoxia.

Methods:Rat renal proximal tubule NRK-52E cells were treated with different concentrations of DFO for 24 hours, followed by evaluation of RTEC injury, using a Cell Counting Kit-8 (CCK-8) to detect cell viability and western blotting to evaluate the expression of transforming growth factor- beta 1 (TGF-β1), α-smooth muscle actin (α-SMA), and hypoxia-inducible factor-1 alpha (HIF-1α) in NRK-52E cells. Then, three groups of NRK-52E cells were used in experiments, including normal control (NC), 25 µM DFO, and 25 µM DFO + MSC-CM. MSC-CM was obtained from the human umbilical cord. MSC-CM was used to culture cells for 12 hours before DFO treatment, then fresh MSC-CM and 25 µM DFO were added, and cells were cultured for another 24 hours before analysis.

Results:Western blotting and cellular immunofluorescence staining showed culture of NRK-52E cells in 25 µM DFO for 24 hours induced HIF-1α and nuclear receptor coactivator-1 (NCoA-1), simulating hypoxia. MSC-CM could inhibit the DFO-induced up-regulation of α-SMA, TGF-β1, HIF-1α and NCoA-1.

Conclusion:Our results suggest that MSC-CM has a protective effect on RTECs by down-regulating HIF-1α and NCoA-1, which may be the harmful factors in renal injury.

About the authors

Chunling Liao

Department of Nephrology, Second Affiliated Hospital of Shantou University Medical College

Email: info@benthamscience.net

Yiping Liu

Department of Nephrology, Second Affiliated Hospital of Shantou University Medical College

Email: info@benthamscience.net

Yongda Lin

Department of Nephrology, Second Affiliated Hospital of Shantou University Medical College

Email: info@benthamscience.net

Jiali Wang

Department of Nephrology, Second Affiliated Hospital of Shantou University Medical College

Email: info@benthamscience.net

Tianbiao Zhou

Department of Nephrology, Second Affiliated Hospital of Shantou University Medical College

Author for correspondence.
Email: info@benthamscience.net

Wenjuan Weng

Department of Nephrology, Second Affiliated Hospital of Shantou University Medical College

Author for correspondence.
Email: info@benthamscience.net

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