Nat Prod Sci.  2018 Jun;24(2):99-102. 10.20307/nps.2018.24.2.99.

Ameliorative Effects of Ombuoside on Dopamine Biosynthesis in PC12 Cells

• Davaasambuu U
• Park KH
• Park HJ
• Choi HS
• Lee CK
• Hwang BY
• Lee MK

• KMID: 2416069
• DOI: http://doi.org/10.20307/nps.2018.24.2.99

Abstract

This study investigated the effects of ombuoside, a flavonol glycoside, on dopamine biosynthesis in PC12 cells. Ombuoside at concentrations of 1, 5, and 10 µM increased intracellular dopamine levels at 1 - 24 h. Ombuoside (1, 5, and 10 µM) also significantly increased the phosphorylation of tyrosine hydroxylase (TH) (Ser40) and cyclic AMP-response element binding protein (CREB) (Ser133) at 0.5 - 6 h. In addition, ombuoside (1, 5, and 10 µM) combined with L-DOPA (20, 100, and 200 µM) further increased intracellular dopamine levels for 24 h compared to L-DOPA alone. These results suggest that ombuoside regulates dopamine biosynthesis by modulating TH and CREB activation in PC12 cells.

Keyword

Ombuoside; Dopamine biosynthesis; TH; CREB; PC12 cells

MeSH Terms

Animals
Carrier Proteins
Dopamine*
Levodopa
PC12 Cells*
Phosphorylation
Tyrosine 3-Monooxygenase
Carrier Proteins
Dopamine
Levodopa
Tyrosine 3-Monooxygenase

Figure

• 

  Fig. 1. Effects of ombuoside on dopamine content in PC12 cells. PC12 cells were treated with ombuoside (1, 5, and 10 µM) and then incubated for 1 − 24 h. PC12 cells were harvested and then dopamine levels were determined by an HPLC method. The mean control levels of intracellular dopamine (0 h) were 3.78 ± 0.31 nmol/mg protein. The results are expressed as the means ± S.E.M. (n = 6 − 8). ∗P < 0.05 compared to the control levels (ANOVA followed by Dunnett's test).

• 

  Fig. 2. Effects of ombuoside on the phosphorylation of TH (Ser40) in PC12 cells. PC12 cells were treated with ombuoside (1, 5, and 10 µM) for 0.5 − 6 h. Immunoblots of lysates of cell pellets were probed with phospho-TH and then the phosphorylation of TH (Ser40) (p-TH) and total TH were analyzed by western blotting. The values of the relative density ratios of p-TH/TH are expressed in arbitrary units. The results are expressed as the means ± S.E.M. (n =4). ∗P<0.05 compared to the control levels (ANOVA followed by Dunnett's test).

• 

  Fig. 3. Effects of ombuoside on the phosphorylation of CREB (Ser133) in PC12 cells. PC12 cells were treated with ombuoside (1, 5, and 10 µM) for 0.5 − 6 h. Immunoblots of lysates of cell pellets were probed with phospho-CREB and then the phosphorylation of CREB (Ser133) (p-CREB) and total CREB were analyzed by western blotting. The values of the relative density ratios of p-CREB/CREB are expressed in arbitrary units. The results are expressed as the means ± S.E.M. (n = 4). ∗P < 0.05 compared to the control levels (ANOVA followed by Dunnett's test).

• 

  Fig. 4. Effects of ombuoside on L-DOPA-induced dopamine levels in PC12 cells. PC12 cells were exposed to L-DOPA (20, 100, and 200 µM) in the absence or presence of ombuoside (1, 5 and 10 µM) for 24 h. PC12 cells were harvested and then dopamine levels were determined by an HPLC method. The mean control levels of intracellular dopamine were 3.56 ± 0.24 nmol/mg protein. The results are expressed as the means ± S.E.M. (n = 6 − 8). ∗P < 0.05 compared to the control levels (untreated cells), #P < 0.05 compared to corresponding L-DOPA treatment (20, 100, and 200 µM) (ANOVA followed by Dunnett's test).

Reference

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