Abstract
We have previously shown that roflupram (ROF) protects against MPP+-induced neuronal damage in models of Parkinson’s disease (PD). Since impaired degradation of α-synuclein (α-syn) is one of the key factors that lead to PD, here we investigated whether and how ROF affects the degradation of α-syn in rotenone (ROT)-induced PD models in vivo and in vitro. We showed that pretreatment with ROF (10 μM) significantly attenuated cell apoptosis and reduced the level of α-syn in ROT-treated SH-SY5Y cells. Furthermore, ROF significantly enhanced the lysosomal function, as evidenced by the increased levels of mature cathepsin D (CTSD) and lysosomal-associated membrane protein 1 (LAMP1) through increasing NAD+/NADH and the expression of sirtuin 1 (SIRT1). Pretreatment with an SIRT1 inhibitor selisistat (SELI, 10 μM) attenuated the neuroprotection of ROF, ROF-reduced expression of α-syn, and ROF-increased expression levels of LAMP1 and mature CTSD. Moreover, inhibition of CTSD by pepstatin A (20 μM) attenuated ROF-reduced expression of α-syn. In vivo study was conducted in mice exposed to ROT (10 mg·kg−1·d−1, i.g.) for 6 weeks; then, ROT-treated mice received ROF (0.5, 1, or 2 mg·kg−1·d−1; i.g.) for four weeks. ROF significantly ameliorated motor deficits, which was accompanied by increased expression levels of tyrosine hydroxylase, SIRT1, mature CTSD, and LAMP1, and a reduced level of α-syn in the substantia nigra pars compacta. Taken together, these results demonstrate that ROF exerts a neuroprotective action and reduces the α-syn level in PD models. The mechanisms underlying ROF neuroprotective effects appear to be associated with NAD+/SIRT1-dependent activation of lysosomal function.
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Acknowledgements
This work was supported by Guangdong Basic and Applied Basic Research Foundation (2021A1515011133), Science and Technology Program of Guangzhou (No. 202002030494), National Natural Science Foundation of China (Nos. 81773698 and 81974501), Key-Area Research and Development Program of Guangdong Province (No. 2018B030334001), the Program for Changjiang Scholars and Innovative Research Team in University (IRT_16R37).
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WLD, JHZ, YQC, JFX, YYQ, NBC, LL and MFL performed research and analyzed data. WLD, HTW and JPX wrote the paper. HTW and JPX conceived and designed research. All authors have given their final approval for the manuscript.
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Dong, Wl., Zhong, Jh., Chen, Yq. et al. Roflupram protects against rotenone-induced neurotoxicity and facilitates α-synuclein degradation in Parkinson’s disease models. Acta Pharmacol Sin 42, 1991–2003 (2021). https://doi.org/10.1038/s41401-021-00768-4
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DOI: https://doi.org/10.1038/s41401-021-00768-4
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