神経変性疾患の細胞モデルを用いた、発症機序と予防法についての研究

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We have been studying the mechanisms underlying cell death caused by alpha-synuclein using our inducible PC12 cell model of Parkinson's disease. We confirmed induction and suppression of alpha-synuclein in the absence and presence of doxycycline, respectively, with immunocytochemistry and western blotting experiments. We observed cytoplasmic inclusions stained for alpha-synuclein and ubiquitin in A53T mutant cell lines. In cell toxicity evaluation, we employed the trypan-blue exclusion assay and DAPI nuclear staining, finding increased apoptotic cell death after neuron-like differentiation and induction of A53T alpha-synuclein expression. Cell death we observed here was in an alpha-synuclein expression dependent manner.We have proposed to elucidate the role for the NF-kappaB pathway in cell death caused by mutant alpha-synuclein. We found that cytoplasmic inclusions in A53T cell lines were positive for IkappaB immunostaining. We tried three NF-kappaB inhibitors including PDTC,DDTC, and SN5O, finding attenuation of A53T alpha-synuclein-caused cell death in the presence of the inhibitors.Our findings suggest that the NF-kappaB pathway may play a role in mutant alpha-synuclein-caused cell death. In addition, the NF-kappaB pathway may be a potential target to treat neural cell death caused by alpha-synuclein.