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Davaasambuu, Park, Park, Choi, Lee, Hwang, and Lee: Ameliorative Effects of Ombuoside on Dopamine Biosynthesis in PC12 Cells
Original Article

Natural Product Sciences 2018;24(2):99-102.

Published online: 20 January 2018

DOI: https://doi.org/10.20307/nps.2018.24.2.99

Ameliorative Effects of Ombuoside on Dopamine Biosynthesis in PC12 Cells

Uchralsaikhan Davaasambuu1, Keun Hong Park1, Hyun Jin Park1, 2, Hyun Sook Choi3, Chong Kil Lee1, 2, Bang Yeon Hwang1, Myung Koo Lee1, 2, ∗

1College of Pharmacy

2Research Center for Bioresource and Health, Chungbuk National University, Cheongju 28160, Korea

3Department of Food and Nutrition, Chungcheong University, Cheongju 28171, Republic of Korea

∗Author for correspondence Myung Koo Lee, College of Pharmacy and Research Center for Bioresource and Health, Chungbuk National University, Cheongju 28160, Korea. Tel: +82-43-261-2822; E-mail: myklee@chungbuk.ac.kr

Received 1 September 2017       Revised 8 February 2018       Accepted 11 February 2018

Copyright © 2018 The Korean Society of Pharmacognosy

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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.

Keywords: Ombuoside, Dopamine biosynthesis, TH, CREB, PC12 cells

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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).
nps-24-99f1.tif
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).
nps-24-99f2.tif
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).
nps-24-99f3.tif
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).
nps-24-99f4.tif
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