Abstract
Introduction DJ‐1 mutation is a causative reason for familial Parkinson's disease (PD). Leucine166Proline (L166P) and C106S are two important DJ‐1 mutations. In this study, we established hydrogen peroxide (H2O2) induced L166P and C106S DJ‐1‐transfected neuroblastoma (SH‐SY5Y) cellular models of PD and investigated the effects of Cistanche extracts and key bioactive compounds, including acteoside, echinacoside, caffeic acid, and Cistanche total glycosides on these two models.MethodsAfter expressing FLAG‐tagged L166P and C106S DJ‐1 plasmids in Escherichia coli, the expressed plasmids were collected, treated with restriction enzyme, and identified using DNA electrophoresis. After purification, the L166P DJ‐1 and C106S DJ‐1 plasmids were separately transfected into SH‐SY5Y cells using liposomes. Transfected SH‐SY5Y cells were detected by western blotting and immunocytochemistry. Cell viability was determined using MTT assay.ResultsBoth western blotting and immunocytochemistry showed that L166P and C106S DJ‐1 were highly expressed in the transfected SH‐SY5Y cells. MTT assays showed that transfection with L166P or C106S DJ‐1 reduced the viability of SH‐SY5Y cells exposed to H2O2, as compared to untransfected SH‐SY5Y cells. In addition, Cistanche extracts and key bioactive compounds, including acteoside, echinacoside, caffeic acid, and Cistanche total glycosides, significantly inhibited the decreases of cell viability caused by H2O2 in L166P and C106S DJ‐1‐transfected SH‐SY5Y cells.ConclusionsThese findings suggest that we successfully established sensitive and stable H2O2 induced L166P DJ‐1‐ and C106S DJ‐1‐transfected SH‐SY5Y cell models of PD and Cistanche extracts may thus be useful for treating PD.
Highlights
DJ‐1 mutation is a causative reason for familial Parkinson's disease (PD)
Extracts, including acteoside, echinacoside, caffeic acid, and Cistanche total glycosides, were added and the cells were incubated for 6 hr before treatment with 0.2 mM H2O2 for an additional 1 hr
After incubation for 1 hr in the presence of graded concentrations of H2O2 (0.1, 0.2, 0.3, 0.4, 0.5, and 1 mM, respectively), the viabilities of SH‐SY5Y cells transfected with L166P or C106S DJ‐1 were dose‐ dependently reduced as compared to untransfected cells (Table 1)
Summary
The main neurodegenerative diseases include Alzheimer's disease, Parkinson's disease (PD), Huntington's disease, and others. Glial cells within the cortex and substantia nigra and striatum are the main found in areas of DJ‐1 protein (Olzmann et al, 2007). In these cells, DJ‐1 plays the following roles, such as oncogene (Nagakubo et al, 1997), transcriptional regulation (Kim et al, 2005; Niki, Takahashi‐ Niki, Taira, Iguchi‐Ariga, & Agria, 2003), antioxidative stress (Taira et al, 2004; Zhou & Freed, 2005), chaperone (Shendelman, Jonason, Marlinat, Leete, & Abeliovich, 2004; Zhou, Zhu, Wioson, Petsko, & Fink, 2006), and protease (Abou‐Sleiman, Healy, Quinn, Lees, & Wood, 2003). To develop a more physiologically relevant cellular model of PD, in this study we established H2O2 induced L166P and C106S DJ‐1‐transfected SH‐SY5Y cells, and investigated the effects of Cistanche extracts and key bioactive compounds, including acteoside, echinacoside, caffeic acid, and Cistanche total glycosides on these two models, for the first time
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