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Protective Effect of Methylene Blue on Neuronal Damage Induced by Okadaic Acid in Differentiated SK-N-SH Neuroblastoma Cells

SI Yingying1, YUAN Yuting2, QIU Lihong2, LÜ Huanhuan3, ZHOU Hui2, XU Bo1, LI Gang1, LI Ji1, WANG Zhenhua1,*   

  1. 1. Center of Mitochondria and Healthy Aging, College of Life Sciences, Yantai University, Yantai 264005, China;
    2. School of Pharmacy, Shihezi University, Shihezi 832005, China; 3. Northwest Institute of Plateau Biology,
    Chinese Academy of Sciences, Xining 810001, China
  • Online:2016-09-15 Published:2016-09-22

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Abstract:

Objective: To investigate the protective effect and underlying mechanism of methylene blue (MB) on neuronal
damage induced by okadaic acid (OA). Methods: The mature neurons differentiated from SK-N-SH neuroblastoma cells
induced by all-trans retinoic acid (ATRA) were exposed to OA to induce synaptic atrophy and cell injury. The SRB assay
was used to measure the effect of MB on the proliferation of differentiated SK-N-SH cells. Giemsa staining was used to
observe cell morphology. The expression of synaptophysin and the phosphorylation of tau protein were detected by Western
blotting. Results: ATRA inhibited the proliferation of SK-N-SH cells in a dose-dependent manner in vitro. The 10 μmol/L
ATRA exposure for 7 days could induce SK-N-SH neuroblastoma cells differentiation into mature neurons with obviously
extended synapse. Treatment with 40 nmol/L OA resulted in synaptic atrophy in differentiated SK-N-SH neuroblastoma
cells. MB inhibited the proliferation of differentiated SK-N-SH cells in a dose-dependent manner in vitro. Moreover,
MB treatment significantly increased the ratio of axon length to cell body length and the expression of synaptophysin
(P < 0.01), while it decreased the phosphorylation of tau protein at Ser262 site (P < 0.01). Conclusion: MB can alleviate
synaptic atrophy induced by OA, which may be due to the down-regulation of tau phosphorylation at Ser262 site.

Key words: methylene blue, differentiated SK-N-SH neuroblastoma cells, all-trans retinoic acid, okadaic acid, tau phosphorylation

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