| Summary: | 碩士 === 中山醫學大學 === 生物化學研究所 === 92 === Recently, it have been shown that rotenone and MPTP, a specific inhibitor of mitochondrial complexⅠ, is a useful tool in animal model of Parkinson’s disease. In this thesis, a human dopaminergic neuronblastoma SH-SY5Y cell line was used to study the effect of exposure to rotenone and MPTP on cell death, activation of stress-induce pathway and aggregation ofα-synuclein, the characteristic protein accumulated in lewy bodies and mimic the pathogenesis of Parkinson’s disease. Moreover it was investigated that the role of antioxidants play in the rotenone- and MPTP-induced α-synuclein aggregattion, oxidative stress and apoptosis. The results showed that both toxins induce a time and dose-dependant decrease in cell survival with IC50 approached values of 0.5 μM after 48hr for rotenone and 1 mM after 24h for MPTP, respectively. Rotenone and MPTP induced apoptosis was demonstrated by using trypan blue exclusion assay, PI staining, DNA fragmentation, Western blotting assay. In addition, it was found that rotenone and MPTP induced apoptosis an increase in ROS production. The Rotenone- and MPTP-induced cell death and caspase 3 activation were reduced by pretreatment with antioxidants (N-acetyl-cystein (10 mM), Baicalein(40 μM), Acetyl-L-Carnitine(0.5 mM) and Lipoic acid (0.1 mM)),indicated a role of ROS in the neurotoxicity. These results suggested that the antioxidants prevent neurons cells from rotenone and MPTP induce apoptosis through decreasing oxidative stress.
|
|---|
