Adaptive changes in autophagy after UPS impairment in Parkinson's disease

作者：Shen, Yu-fei; Tang, Yu; Zhang, Xiao-jie; Huang, Kai-xing; Le, Wei-dong^*

来源：Acta Pharmacologica Sinica, 2013, 34(5): 667-673.

DOI：10.1038/aps.2012.203

• proteasome inhibitor
• lactacystin
• ubiquitin-proteasome system
• dopaminergic neuron
• Parkinson's disease
• autophagylysosomal pathway
• autophagosome
• mTOR
• Beclin 1

Aim: Ubiquitin-proteasome system (UPS) and autophagosome-lysosome pathway (ALP) are the most important machineries responsible for protein degradation in Parkinson's disease (PD). The aim of this study is to investigate the adaptive alterations in autophagy upon proteasome inhibition in dopaminergic neurons in vitro and in vivo.
Methods: Human dopaminergic neuroblastoma SH-SY5Y cells were treated with the proteasome inhibitor lactacystin (5 mu mol/L) for 5, 12, or 24 h. The expression of autophagy-related proteins in the cells was detected with immunoblotting. UPS-impaired mouse model of PD was established by microinjection of lactacystin (2 mu g) into the left hemisphere of C57BL/6 mice that were sacrificed 2 or 4 weeks later. The midbrain tissues were dissected to assess alterations in autophagy using immunofluorescence, immunoblotting and electron microscopy assays.
Results: Both in SH-SY5Y cells and in the midbrain of UPS-impaired mouse model of PD, treatment with lactacystin significantly increased the expression levels of LC3-I/II and Beclin 1, and reduced the levels of p-mTOR, mTOR and p62/SQSTM1. Furthermore, lactacystin treatment in UPS-impaired mouse model of PD caused significant loss of TH-positive neurons in the substantia nigra, and dramatically increased the number of autophagosomes in the left TH-positive neurons.
Conclusion: Inhibition of UPS by lactacystin in dopaminergic neurons activates another protein degradation system, the ALP, which includes both the mTOR signaling pathway and Beclin 1-associated pathway.

• 出版日期2013-5
• 单位上海市闵行区中心医院; 中国科学院上海生命科学研究院; 上海交通大学