Background/Aims: Cisplatin-induced acute kidney injury (AKI) involves damage to tubular cells via excess reactive oxygen species (ROS) generation. Stem cell-based therapies have shown great promise in AKI treatment. In this study, we aimed to assess the protective effect and mechanism of bone marrow mesenchymal stem cell (BMSC)-derived conditioned medium (CM) against cisplatin-induced AKI. Methods: In vitro, NRK-52E cells were incubated with cisplatin in the presence or absence of CM, followed by the assessment of cell viability, apoptosis and cell cycle distribution. Then, ICG-001 and IWR-1 were used to inhibit the wnt/beta-catenin pathway. Furthermore, intracellular and mitochondrial ROS levels were evaluated using DCFH-DA and MitoSOX, respectively. In vivo, after cisplatin injection, rats were intravenously injected with CM or BMSCs. Sera and kidney tissues were collected on day 3 after cisplatin injection to evaluate changes in renal function and histology. Western blotting and qRT-PCR were employed to determine the expression of wnt/beta-catenin pathway-related genes and proteins. Immunohistochemical staining was used to evaluate tubular beta-catenin expression in kidney biopsy from AKI patients. Results: CM protected NRK-52E cells from cisplatin-induced injury by restoring the wnt4/beta-catenin pathway. In response to ICG-001 and IWR-1, the protective effect of CM was attenuated, characterized by a decrease in cell proliferation and an increase in cell apoptosis and intracellular and mitochondrial ROS levels. Knockdown of beta-catenin using siRNAs also suppressed the mitochondrial biogenesis regulators PGC-1 alpha, TFAM and NRF-1. In the rat model, CM significantly alleviated renal function and histology associated with tubular injury and upregulated wnt4 and beta-catenin. However, the renoprotective effect of CM was blocked by ICG-001, characterized by exacerbated renal function, suppressed PGC-1 alpha expression and increased mitochondrial ROS. Clinical data showed that the tubular beta-catenin level was lower in AKI patients experiencing partial recovery than in patients experiencing complete recovery. Conclusion: The activation of the wnt/beta-catenin pathway by CM protects against cisplatin-induced kidney injury, resulting in reduced apoptosis and intracellular ROS levels. (c) 2017 The Author(s) Published by S. Karger AG, Basel
[1]Fudan Univ, Zhongshan Hosp, Dept Nephrol, 180 Fenglin Rd, Shanghai 200032, Peoples R China.
[2]Shanghai Med Ctr Kidney, Shanghai, Peoples R China.
[3]Shanghai Key Lab Kidney & Blood Purificat, Shanghai, Peoples R China.
[4]Shanghai Inst Kidney & Dialysis, Shanghai, Peoples R China.
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