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1,25-dyhydroxyvitamin D-3 Attenuates L-DOPA-Induced Neurotoxicity in Neural Stem Cells

Authors
Jang, WooyoungPark, Hyun-HeeLee, Kyu-YongLee, Young JooKim, Hee-TaeKoh, Seong-Ho
Issue Date
Apr-2015
Publisher
SPRINGER
Keywords
L-DOPA; Neural stem cells; 1,25-dyhydroxyvitamin D-3; Neuroprotection
Citation
MOLECULAR NEUROBIOLOGY, v.51, no.2, pp.558 - 570
Indexed
SCIE
SCOPUS
Journal Title
MOLECULAR NEUROBIOLOGY
Volume
51
Number
2
Start Page
558
End Page
570
URI
https://scholarworks.bwise.kr/hanyang/handle/2021.sw.hanyang/157621
DOI
10.1007/s12035-014-8835-1
ISSN
0893-7648
Abstract
The neurotoxicity of levodopa (l-DOPA) on neural stem cells (NSCs) and treatment strategies to protect NSCs from this neurotoxicity remain to be elucidated. Recently, an active form of vitamin D-3 has been reported to display neuroprotective properties. Therefore, we investigated the protective effect of 1,25-dyhydroxyvitamin D-3 (calcitriol) on l-DOPA-induced NSC injury. We measured cell viability via the cell counting kit-8 (CCK-8) and lactate dehydrogenase (LDH) assays and Annexin V/PI staining followed by flow cytometry, cell proliferation using the BrdU and colony-forming unit (CFU) assays, cell differentiation via immunocytochemistry, the levels of free radicals via 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA) staining, apoptosis via DAPI and TUNEL staining, and intracellular signaling protein expression via Western blot. Antibody microarrays were also employed to detect changes in the expression of prosurvival- and death-related proteins. Treatment of NSCs with l-DOPA reduced their viability and proliferation. This treatment also increased the levels of free radicals and decreased the expression levels of intracellular signaling proteins that are associated with cell survival. However, simultaneous exposure to calcitriol significantly reduced these effects. The calcitriol-mediated protection against l-DOPA toxicity was blocked by the phosphoinositide 3-kinase (PI3K) inhibitor LY294004. l-DOPA also inhibited the expression of Nestin and Ki-67, and co-treatment with calcitriol alleviated these effects. The expression levels of GFAP, DCX, and Tuj1 were not significantly affected by treatment with l-DOPA or calcitriol. Calcitriol protects against l-DOPA-induced NSC injury by promoting prosurvival signaling, including activation of the PI3K pathway, and reducing oxidative stress.
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