Autophagy protects the rotenone-induced cell death in alpha-synuclein overexpressing SH-SY5Y cells.
ABSTRACT Loss of dopaminergic cells induced by alpha-synuclein accumulation in substantia nigra causes the development of Parkinson's disease (PD). To date, although autophagy has been implicated in the pathology of PD, the molecular mechanism is still unclear. To study the role of autophagy in PD pathogenesis, we established stable SH-SY5Y cell lines overexpressing wild-type or mutant alpha-synuclein proteins (A30P or A53T). Overexpression of mutant alpha-synuclein induced some protein aggregates and cell death in the absence of drug. LC3-II protein, a critical marker for autophagy, was produced in an autophagy-dependent manner. The rotenone-induced cell death was interrupted by autophagy stimulation. Autophagy activation also restored the mitochondrial membrane potential (MMP) impaired by rotenone in mutant alpha-synuclein expressing cells. Additionally, autophagy activation significantly relieved rotenone-induced ROS accumulation and HIF-1alpha expression in neuronal cells expressing mutant alpha-synuclein proteins. These findings indicate that autophagy plays an important scavenger role against harmful influence of toxic protein aggregates produced in rotenone-treated cells.
SourceAvailable from: Carsten Culmsee[Show abstract] [Hide abstract]
ABSTRACT: Parkinson's disease (PD) is the most frequent neurodegenerative movement disorder. Presently, there is no causal therapy available to slow down or halt disease progression. The presynaptic protein alpha-synuclein aggregates to form intraneuronal Lewy bodies in PD. It is generally believed that intermediates on the way from monomers to the large aggregates would mediate neurotoxicity, but the precise species and mechanism responsible for neuronal death are controversially debated. To study alpha-synuclein-mediated toxicity, we developed a new model in which moderate overexpression of wild-type alpha-synuclein led to gradual death of human postmitotic dopaminergic neurons. In accordance with findings in postmortem PD brains, small oligomeric species occurred and the autophagic flux was impaired in our model. The phenothiazine neuroleptic trifluoperazine, an activator of macroautophagy, selectively reduced one particular alpha-synuclein species and rescued cells. Inversely, blocking of autophagy led to an accumulation of this oligomeric species and increased cell death. These data show that activation of autophagy is a promising approach to protect against alpha-synuclein pathology and likely acts by targeting one specific alpha-synuclein species.Neurobiology of aging 01/2014; 35(7). DOI:10.1016/j.neurobiolaging.2014.01.027 · 4.85 Impact Factor
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ABSTRACT: Objectives Modified Yeolda-Hanso tang (MYH) is a traditional herbal formula in Korea for various diseases. MYH is containing the 10 herbs : Pueraria lobata (Willd.) Ohwi, Angelica tenuissima Nakai, Scutellaria baicalensis Georgi, Platycodon grandiflorum (Jacq), Angelicae Dahurica, Cimicifuga heracleifolia Kom, Raphanus sativa L., Polygala tenuifolia (Willd), Acorus gramineus Soland and Dimocarpus longan Lour. The 10 herbs is constituted as a ratio of the 6:4:2:1:2:2:2:4:6:6. We investigated neuroprotective effects of MYH on human neuroblastoma SH-SY5Y cells and evaluated the ability of MYH to prevent and treat for neurodegenerative diseases such as Parkinson's disease via basal autophagy enhancement. Methods Pharmacological induction of Autophagy by MYH in SH-SY5Y cells: Induction of autophagy by MYH in human neuroblastoma SH-SY5Y cells was carreid out by immunoblot analysis with several autophagy markers. SH-SY5Y cells were treated with MYH at the concentration of 400 and for 24 hr. Specifically, the autophagosome proteins LC3 II and Atg5 levels were increased and autophagy pathway related proteins such as beclin-1, PI3 Kinase class III protein, ULK1, mTOR and AMPK were activated. Conclusions MYH can enhance the induction of autophagy through key regulator AMPK, mTOR, and Beclin-1 and it should be considered as a possible candidate of neuroprotective agents for such as Parkinson's disease.09/2013; 25(3). DOI:10.7730/JSCM.2013.25.3.208
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ABSTRACT: Autophagy is a series of catabolic process mediating the bulk degradation of intracellular proteins and organelles through formation of a double-membrane vesicle, known as an autophagosome, and fusing with lysosome. Autophagy plays an important role of death-survival decisions in neuronal cells, which may influence to several neurodegenerative disorders including Parkinson's disease. Chebulagic acid, the major constituent of Terminalia chebula and Phyllanthus emblica, is a benzopyran tannin compound with various kinds of beneficial effects. This study was performed to investigate the autophagy enhancing effect of chebulagic acid on human neuroblastoma SH-SY5Y cell lines. We determined the effect of chebulagic acid on expression levels of autophago-some marker proteins such as, DOR/TP53INP2, Golgi-associated ATPase Enhancer of 16 kDa (GATE 16) and Light chain 3 II (LC3 II), as well as those of its upstream pathway proteins, AMP-activated protein kinase (AMPK), mammalian target of rapamycin (mTOR) and Beclin-1. All of those proteins were modulated by chebulagic acid treatment in a way of enhancing the autophagy. Additionally in our study, chebulagic acid also showed a protective effect against 1-methyl-4-phenylpyridinium (MPP(+)) - induced cytotoxicity which mimics the pathological symptom of Parkinson's disease. This effect seems partially mediated by enhanced autophagy which increased the degradation of aggregated or misfolded proteins from cells. This study suggests that chebulagic acid is an attractive candidate as an autophagy-enhancing agent and therefore, it may provide a promising strategy to prevent or cure the diseases caused by accumulation of abnormal proteins including Parkinson's disease.Biomolecules and Therapeutics 07/2014; 22(4):275-81. DOI:10.4062/biomolther.2014.068 · 0.84 Impact Factor