Abstract
Activation of pro-inflammatory microglia is an important mechanism of the cerebral ischemia–reperfusion (I/R)-induced neuronal injury and dysfunction. Mesenchymal stem cells (MSCs) together with their paracrine factors demonstrated curative potential in immune disorders and inflammatory diseases, as well as in ischemic diseases. However, it remains unclear whether conditioned medium from MSCs could effectively regulate the activation and polarization of microglia exposed to I/R stimulation. In this study, we investigated the effects of conditioned medium from bone marrow MSCs (BMSCs-CM) on I/R-stimulated microglia and the potential mechanism involved, as well as the way to obtain more effective BMSCs-CM. First, cell model of oxygen–glucose deprivation/reoxygenation (OGD/R) was established in microglia to mimic the I/R. BMSCs-CM from different culture conditions (normoxic: 21% O2; hypoxic: 1% O2; hypoxia preconditioning: preconditioning with 1% O2 for 24 h) was used to treat the microglia. Our results showed that BMSCs-CM effectively promoted the survival and alleviated the injury of microglia. Moreover, in microglia exposed to OGD/R, BMSCs-CM inhibited significantly the expression of pro-inflammatory cytokines (TNF-α, IL-1β, and IL-6), CD86 and inducible nitric oxide synthase, whereas upregulated the levels of anti-inflammatory cytokine (IL-10), CD206 and Arginase-1. These results suggested that BMSCs-CM promoted the polarization of anti-inflammatory microglia. In particular, BMSCs-CM from cultures with hypoxia preconditioning was more effective in alleviating cell injury and promoting anti-inflammatory microglia polarization than BMSCs-CM from normoxic cultures and from hypoxic cultures. Furthermore, inhibition of exosomes secretion could largely mitigate these effects of BMSCs-CM. In conclusion, our results suggested that hypoxia preconditioning of BMSCs could enhance the efficacy of BMSCs-CM in alleviating OGD/R-induced injury and in promoting the anti-inflammatory polarization of microglia, and these beneficial effects of BMSCs-CM owed substantially to exosomes.
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This work was supported by the National Natural Science Foundation of China (Grant No. 81771707), and the Fundamental Research Funds for the Central Universities of China (Grant No. 2042018kf0235). The funders had no role in the study design, the data collection and analysis or the decision to publish.
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YL and HY performed interpretation and drafted the manuscript. YL, YC, XH, YL and HY contributed to designing experiments and revising the manuscript. HY and ZX performed most of the experiments and data analysis. GQ performed the experiments of BV2 microglia culture and CCK-8 test. LL and HW performed culture and characterization of BMSCs. XL and XX performed the LDH and ROS analysis. YL and YL provided the experimental reagents and materials. All authors have read the manuscript and approved the final manuscript.
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All experimental animal procedures were approved by the Ethics Committee on Animal Care and Use of Wuhan University, Wuhan, China (Permit Number 20170303).
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Yu, H., Xu, Z., Qu, G. et al. Hypoxic Preconditioning Enhances the Efficacy of Mesenchymal Stem Cells-Derived Conditioned Medium in Switching Microglia toward Anti-inflammatory Polarization in Ischemia/Reperfusion. Cell Mol Neurobiol 41, 505–524 (2021). https://doi.org/10.1007/s10571-020-00868-5
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DOI: https://doi.org/10.1007/s10571-020-00868-5