Abstract
We have previously shown that PPAR-γ agonist 15d-PGJ2 inhibited neuronal autophagy after cerebral ischemia/reperfusion injury. However, the underlying mechanism of its regulatory role in neuronal autophagy remains unclear. This study was designed to test the hypothesis that 15d-PGJ2 upregulated Bcl-2 which binds to Beclin 1, and thereby inhibits autophagy. We performed cell viability assay, cytotoxicity assay, western blot, and co-immunoprecipitation to analyze autophagy activities in vitro model of oxygen–glucose deprivation/reoxygenation (OGD/R). OGD/R induced autophagy in cultured cortical neurons. 15d-PGJ2 treatment significantly decreased LC3-II/LC3-I ratio and Beclin 1 expression, but increased p62 expression. Autophagic inhibitor 3-methyladenine decreased LC3-II levels, increased neuronal cell viability, and mimicked some protective effect of 15d-PGJ2 against OGD/R injury. OGD/R-induced autophagy coincided with decreases in Bcl-2 expression and increases in Beclin 1 expression. 15d-PGJ2 treatment upregulated Bcl-2 expression and decreased Beclin 1 expression, and inhibit the dissociation of Beclin1 from Bcl-2 significantly. Bcl-2 siRNA abrogated the effect of 15d-PGJ2 on Beclin 1, LC3-II and p62, and influence cell viability and LDH level, while scRNA did not. PPAR-γ agonist 15d-PGJ2 exerts neuroprotection partially via inhibiting neuronal autophagy after OGD/R injury. The inhibition of autophagy by 15d-PGJ2 is mediated through upregulation of Bcl-2.
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Acknowledgments
This work was supported by National Natural Science Foundation of China 81000488 (F.X.), Fudan University Young Teacher Capability Enhancement Program 20520133268 (F.X.), and Nanjing Medical Science and Technology Development Program YKK11124 (H.D.Q.).
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The authors have declared that no competing interests exist.
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Haidong Qing, Weiguo Tan, Feng Xu and Zizheng Wang contributed equally to this work.
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Qin, H., Tan, W., Zhang, Z. et al. 15d-Prostaglandin J2 Protects Cortical Neurons Against Oxygen–Glucose Deprivation/Reoxygenation Injury: Involvement of Inhibiting Autophagy Through Upregulation of Bcl-2. Cell Mol Neurobiol 35, 303–312 (2015). https://doi.org/10.1007/s10571-014-0125-y
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DOI: https://doi.org/10.1007/s10571-014-0125-y