Abstract
Intracerebral hemorrhage (ICH) is a devastating disease, in which neuroinflammation substantially contributes to brain injury. Uncoupling protein 2 (UCP2) is a member of the mitochondrial anion carrier family, which uncouples oxidative phosphorylation from ATP synthesis by facilitating proton leak across the mitochondrial inner membrane. UCP2 has been reported to modulate inflammation. In this study we investigated whether and how UCP2 modulated neuroinflammation through microglia/macrophages following ICH in vitro and in vivo. We used an in vitro neuroinflammation model in murine BV2 microglia to mimic microglial activation following ICH. ICH in vivo model was established in mice through collagenase infusion into the left striatum. ICH mice were treated with anetholetrithione (ADT, 50 mg· kg−1 ·d−1, ip) or the classical protonophoric uncoupler FCCP (injected into hemorrhagic striatum). We showed that the expression and mitochondrial location of microglial UCP2 were not changed in both in vitro and in vivo ICH models. Knockdown of UCP2 exacerbated neuroinflammation in BV2 microglia and mouse ICH models, suggesting that endogenous UCP2 inhibited neuroinflammation and therefore played a protective role following ICH. ADT enhanced mitochondrial ROS production thus inducing mitochondrial uncoupling and activating UCP2 in microglia. ADT robustly suppressed neuroinflammation, attenuated brain edema and improved neurological deficits following ICH, and these effects were countered by striatal knockdown of UCP2. ADT enhanced AMP-activated protein kinase (AMPK) activation in the hemorrhagic brain, which was abrogated by striatal knockdown of UCP2. Moreover, striatal knockdown of AMPK abolished the suppression of neuroinflammation by ADT following ICH. On the other hand, FCCP-induced mitochondrial uncoupling was independent of UCP2 in microglia; and striatal knockdown of UCP2 did not abrogate the suppression of neuroinflammation by FCCP in ICH mice. In conclusion, the uncoupling activity is essential for suppression of neuroinflammation by UCP2. We prove for the first time the concept that activators of endogenous UCP2 such as anetholetrithione are a new class of uncouplers with translational significance.
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Acknowledgements
This work was supported by following grants: National Natural Science Foundation of China (81971119, 82071469, 81571124, 81671310), Priority Academic Program Development of the Jiangsu Higher Education Institutions (PAPD), Suzhou Clinical Research Center of Neurological Disease (Szzx201503) and the Jiangsu key laboratory grant (BM2013003).
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JJ, JC, and ZCH conceived the concept, designed the research, interpreted data, and wrote the paper. XLY, FYX and JJJ carried out the cell culture, Western blotting, Histochemistry, animal model and behavior tests. PS, YQP, MJH, ZCW and SJY participated in the above work. XLY, FYX and JJJ acquired and analyzed data. FYX drafted all figures. All authors read and approved the final paper.
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Yan, Xl., Xu, Fy., Ji, Jj. et al. Activation of UCP2 by anethole trithione suppresses neuroinflammation after intracerebral hemorrhage. Acta Pharmacol Sin 43, 811–828 (2022). https://doi.org/10.1038/s41401-021-00698-1
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DOI: https://doi.org/10.1038/s41401-021-00698-1
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