摘要：

Objective- Establish a cellular model for studying the Aβ-mediated mitochondrial dysfunction and neuronal stress in Alzheimer's disease. Methods- The full length cDNA fragment of cyclophilin D (CCDS7358.1) was composed after codon optimization, then amplified by PCR and ligated into pcDNA3.1(+)-myc-his vector to construct the recombinant plasmid pcDNA3.1-myc-his-CypD. The SH-SY5Y neuroblastoma cells were transfected with pcDNA3.1-myc-his-CypD using lipofectamine2000 reagent, and stable transfectants were selected in 10000μg/mL G418. Transfection efficiency was examined with western bolt. Results- The sequencing analysis confirmed the accuracy of the eukaryotic expression vector pcDNA3.1-myc-his-CypD. There is no obvious effect of CypD overexpression on cell morphology, but the cell growth rate reduced. Conclusion- The establishment of stable SH-SY5Y CypD cell line provided a cellular model for mitochondrial related pathophysiology and therapy research in Alzheimer's disease.

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