Title:Mesenchymal Stem Cell-conditioned Medium Protecting Renal Tubular Epithelial Cells by Inhibiting Hypoxia-inducible Factor-1α and Nuclear Receptor Coactivator-1
Volume: 19
Issue: 10
Author(s): Chunling Liao, Yiping Liu, Yongda Lin, Jiali Wang, Tianbiao Zhou*Wenjuan Weng*
Affiliation:
- Department of Nephrology, the Second Affiliated Hospital of Shantou University Medical College, Shantou, 515041,
China
- Department of Nephrology, the Second Affiliated Hospital of Shantou University Medical College, Shantou, 515041,
China
Keywords:
Acute kidney injury (AKI), renal tubular epithelial cells (RTECs), deferoxamine (DFO), MSC-conditioned medium (MSC-CM), HIF-1α, NCoA-1.
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.
