Journal List > Nat Prod Sci > v.23(3) > 1060659

TOOLS
Park, Lee, Kim, Jang, Ha, Oh, and Park: Reserpine treatment activates AMP activated protein kinase (AMPK)
Original Article

Natural Product Sciences 2017;23(3):157-161.

Published online: 20 January 2017

DOI: https://doi.org/10.20307/nps.2017.23.3.157

Reserpine treatment activates AMP activated protein kinase (AMPK)

Rackhyun Park1, †, Kang II Lee1, †, Hyunju Kim1, Minsu Jang1, Thi Kim Quy Ha2, Won Keun Oh2, Junsoo Park1, ∗

1Division of Biological Science and Technology, Yonsei University, Wonju 26493, Republic of Korea

2Korea Bioactive Natural Material Bank, College of Pharmacy, Seoul National University, Seoul 08826, Republic of Korea

∗Author for correspondence Junsoo Park, Ph.D, Yonsei University, Division of Biological Sciences and Technology 1 Yonseidae-gil, Wonju City, Kangwon province, 26493, Korea. Tel: +82-33-760-2560; Fax: +82-33-760-2183; E-mail: junsoo@yonsei.ac.kr

† These two authors contributed equally to this work

Received 18 January 2017       Revised 4 May 2017       Accepted 4 May 2017

Copyright © 2017 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

Reserpine is a well-known medicine for the treatment of hypertension, however the role of reserpine in cell signaling is not fully understood. Here, we report that reserpine treatment induces the phosphorylation of AMP activated protein kinase (AMPK) at threonine 172 (T172) in PC12 cells. Phosphorylation of AMPK T172 is regulated by upstream signaling molecules, and the increase of phospho-T172 indicates that AMPK is activated. When we examined the FOXO3a dependent transcription by using the FHRE-Luc reporter assay, reserpine treatment repressed the FHRE-Luc reporter activity in a dose dependent manner. Finally, we showed that reserpine treatment induced the phosphorylation of AMPK as well as cell death in MCF-7 cells. These results suggest that AMPK is a potential cellular target of reserpine.

Keywords: Reserpine, AMPK, FOXO3a, PC12, MCF-7

References

(1). Hardie D. G.., Schaffer B. E.., Brunet A.Trends Cell Biol. 2016. 26:190–201.
(2). Shaw R. J.., Kosmatka M.., Bardeesy N.., Hurley R. L.., Witters L. A.., DePinho R. A.., Cantley L. C. Proc. Natl. Acad. Sci. U. S.A. 2004. 101:3329–3335.
(3). Hardie D. G.., Scott J. W.., Pan D. A.., Hudson E. R.FEBS Lett. 2003. 546:113–120.
crossref
(4). Kemp B. E.., Mitchelhill K. I.., Stapleton D.., Michell B. J.., Chen Z. P.., Witters L. A.Trends Biochem. Sci. 1999. 24:22–25.
(5). Adams J.., Chen Z. P.., Van Denderen B. J.., Morton C. J.., Parker M. W.., Witters L. A.., Stapleton D.., Kemp B. E.Protein Sci. 2004. 13:155–165.
(6). Woods A.., Vertommen D.., Neumann D.., Turk R.., Bayliss J.., Schlattner U.., Wallimann T.., Carling D.., Rider M. H. J.Biol. Chem. 2003. 278:28434–28442.
(7). Li N.., Huang D.., Lu N.., Luo L.Oncol. Rep. 2015. 34:2821–2826.
(8). Ben Sahra I.., Laurent K.., Loubat A.., Giorgetti-Peraldi S.., Colosetti P.., Auberger P.., Tanti J. F.., Le Marchand-Brustel Y.., Bost F.Oncogene. 2008. 27:3576–3586.
(9). Ben Sahra I.., Le Marchand-Brustel Y.., Tanti J. F.., Bost F.Mol. Cancer Ther. 2010. 9:1092–1099.
(10). Plummer A. J.., Earl A.., Schneider J. A.., Trapold J.., Barrett W.Ann. N Y Acad. Sci. 1954. 59:8–21.
(11). López-Muñoz F.., Bhatara V. S.., Alamo C.., Cuenca E.Actas Esp. Psiquiatr. 2004. 32:387–395.
(12). Guo Z.., Liu X.., Huang H.PLoS One. 2015. 10:, e0138619.
(13). Eiden L. E.., Weihe E.Ann. N Y Acad. Sci. 2011. 1216:86–98.
(14). Leão A. H.., Sarmento-Silva A. J.., Santos J. R.., Ribeiro A. M.., Silva R. H.Brain Pathol. 2015. 25:377–390.
(15). Fernandes V. S.., Santos J. R.., Leão A. H.., Medeiros A. M.., Melo T. G.., Izídio G. S.., Cabral A.., Ribeiro R. A.., Abílio V. C.., Ribeiro A. M.., Silva R. H.Behav Brain Res. 2012. 231:154–163.
(16). Lee K. I.., Kim M. J.., Koh H.., Lee J. I.., Namkoong S.., Oh W. K.., Park J.Biochem. Biophys. Res. Commun. 2015. 462:402–408.
(17). Brunet A.., Bonni A.., Zigmond M. J.., Lin M. Z.., Juo P.., Hu L. S.., Anderson M. J.., Arden K. C.., Blenis J.., Greenberg M. E.Cell. 1999. 96:857–868.
(18). Takayama H.., Misu H.., Iwama H.., Chikamoto K.., Saito Y.., Murao K.., Teraguchi A.., Lan F.., Kikuchi A.., Saito R.., Tajima N.., Shirasaki T.., Matsugo S.., Miyamoto K.., Kaneko S.., Takamura T. J.Biol. Chem. 2014. 289:335–345.
(19). Takayama H.., Misu H.., Iwama H.., Chikamoto K.., Raynaud S.., Auberger P.Cancer Res. 2010. 70:1042–1052.

Fig. 1.
Reserpine activates AMPK in PC12 cells. (a) Chemical structure of reserpine (b) Reserpine treatment induced the phosphorylation of AMPK. PC12 cells were incubated with various concentrations of reserpine (0, 1, 2.5, 5 and 10 µM) for 24 h, and the cell lysates were subjected to Western blotting with phospho-AMPK (T172) or AMPK antibodies. Experiments were repeated three times with similar observations, and representative data is shown. The levels of phospho-AMPK/AMPK were analyzed. (c) Compound C treatment inhibited the activation of AMPK induced by reserpine treatment. PC12 cells were incubated with reserpine for 24 h in the presence or absence of Compound C.
nps-23-157f1.tif
Fig. 2.
Reserpine induces cell death in PC12 cells. (a) Reserpine induces the cell death. PC12 cells were incubated with reserpine for 24 h and 48 h, and cell viability was measured using the MTT assay in quadruplicate. The graph shows the average and the standard deviation (SD). Control vs. reserpine treatment, ∗P < 0.05, ∗∗P < 0.005, ∗∗∗P < 0.0005. (b) Reserpine induces apoptosis. PC12 cells were treated with reserpine (10 µM) for 24 h. The cells were stained with propidium iodide and analyzed using flow cytometry for DNA content.
nps-23-157f2.tif
Fig. 3.
Reserpine treatment deregulates the FHRE-Luc reporter activity. HEK293T cells were transfected with FHRE-Luc reporter plasmid. Twenty four h after transfection, cells were treated with various concentrations of reserpine (0, 1, 2.5, 5 and 10 µM) for 24 h and luciferase activity was measured. The luciferase activity was normalized to renilla luciferase activity and the experiment was performed in quadruplicate. The graph shows the average and the standard deviation (SD). Control vs. reserpine treatment, ∗P <0.005, ∗∗P <0.001.
nps-23-157f3.tif
Fig. 4.
Reserpine treatment activates AMPK in MCF-7 cells and induces cell death. (a) MCF-7 cells were incubated with reserpine for 24 h, and the levels of phosphor-AMPK / AMPK in MCF-7 were analyzed. Experiments were repeated three times with similar observations, and representative data is shown. (b) Reserpine treatment induces the cell death. MCF-7 cells were incubated with reserpine for 24 h and 48 h, and cell viability was measured using the MTT assay in quadruplicate. The graph shows the average and the standard deviation (SD). Control vs. reserpine treatment, ∗P < 0.005, ∗∗P < 0.0005. (b) Reserpine increases sub-G1 population. MCF-7 cells were treated with reserpine (10 µM) for 24 h. The cells were stained with propidium iodide and analyzed using flow cytometry for DNA content.
nps-23-157f4.tif
TOOLS
Similar articles