Nitric oxide prevents 6-hydroxydopamine-induced apoptosis in PC12 cells through cGMP-dependent PI3 kinase/Akt activation

Kwon Soo Ha, Ki Mo Kim, Young Guen Kwon, Se Kyung Bai, Woo Dong Nam, Young Min Yoo, Peter K.M. Kim, Hun Taeg Chung, Timothy R. Billiar, Young Myeong Kim

Research output: Contribution to journalArticle

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Abstract

Nitric oxide (NO) functions not only as an important signaling molecule in the brain by producing cGMP, but also regulates neuronal cell apoptosis. The mechanism by which NO regulates apoptosis is unclear. In this study, we demonstrated that NO, produced either from the NO donor S-nitroso-N-acetyl-D,L-penicillamine (SNAP) or by transfection of neuronal NO synthase, suppressed 6-hydroxydopamine (6-OHDA)-induced apoptosis in PC12 cells by inhibiting mitochondrial cytochrome c release, caspase-3 and -9 activation, and DNA fragmentation. This protection was significantly reversed by the soluble guanylyl cyclase inhibitor 1H-(1,2,4)-oxadiazole[4,3-a]quinoxalon-1-one, indicating that cGMP is a key mediator in NO-mediated anti-apoptosis. Moreover, the membrane-permeable cGMP analog 8-Br-cGMP inhibited 6-OHDA-induced apoptosis. These anti-apoptotic effects of SNAP and 8-Br-cGMP were suppressed by cGMP-dependent protein kinase G (PKG) inhibitor KT5823, indicating that PKG is a downstream signal mediator in the suppression of apoptosis by NO and cGMP. Both SNAP and 8-Br-cGMP induced endogenous Akt activation and Bad phosphorylation, resulting in the inhibition of Bad translocation to mitochondria; these effects were inhibited by KT5823 and the phosphatidylinositol 3-kinase (PI3K) inhibitors LY294002 and Wortmannin. Our data suggest that the NO/cGMP pathway suppresses 6-OHDA-induced PC 12 cell apoptosis by suppressing the mitochondrial apoptosis signal via PKG/PI3K/Akt-dependent Bad phosphorylation. - Ha, K-S., Kim, K. M., Kwon, Y.-G., Bai, S.-K., Nam, W.-D., Yoo, Y.-M., Kim, P. K. M., Chung, H.-T., Billiar, T. R., Kim, Y.-M. Nitric oxide prevents 6-hydroxydopamine-induced apoptosis in PC12 cells through cGMP-dependent PI3 kinase/Akt activation.

Original languageEnglish
Pages (from-to)1036-1047
Number of pages12
JournalFASEB Journal
Volume17
Issue number9
DOIs
Publication statusPublished - 2003 Jun 1

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Oxidopamine
PC12 Cells
Phosphatidylinositol 3-Kinases
Nitric Oxide
Chemical activation
Apoptosis
Cyclic GMP-Dependent Protein Kinases
Penicillamine
Phosphatidylinositol 3-Kinase
Phosphorylation
Oxadiazoles
Nitric Oxide Synthase Type I
2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one
Mitochondria
Nitric Oxide Donors
Caspase 9
Guanylate Cyclase
DNA Fragmentation
Protein Kinase Inhibitors
Cytochromes c

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Genetics

Cite this

Ha, Kwon Soo ; Kim, Ki Mo ; Kwon, Young Guen ; Bai, Se Kyung ; Nam, Woo Dong ; Yoo, Young Min ; Kim, Peter K.M. ; Chung, Hun Taeg ; Billiar, Timothy R. ; Kim, Young Myeong. / Nitric oxide prevents 6-hydroxydopamine-induced apoptosis in PC12 cells through cGMP-dependent PI3 kinase/Akt activation. In: FASEB Journal. 2003 ; Vol. 17, No. 9. pp. 1036-1047.
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abstract = "Nitric oxide (NO) functions not only as an important signaling molecule in the brain by producing cGMP, but also regulates neuronal cell apoptosis. The mechanism by which NO regulates apoptosis is unclear. In this study, we demonstrated that NO, produced either from the NO donor S-nitroso-N-acetyl-D,L-penicillamine (SNAP) or by transfection of neuronal NO synthase, suppressed 6-hydroxydopamine (6-OHDA)-induced apoptosis in PC12 cells by inhibiting mitochondrial cytochrome c release, caspase-3 and -9 activation, and DNA fragmentation. This protection was significantly reversed by the soluble guanylyl cyclase inhibitor 1H-(1,2,4)-oxadiazole[4,3-a]quinoxalon-1-one, indicating that cGMP is a key mediator in NO-mediated anti-apoptosis. Moreover, the membrane-permeable cGMP analog 8-Br-cGMP inhibited 6-OHDA-induced apoptosis. These anti-apoptotic effects of SNAP and 8-Br-cGMP were suppressed by cGMP-dependent protein kinase G (PKG) inhibitor KT5823, indicating that PKG is a downstream signal mediator in the suppression of apoptosis by NO and cGMP. Both SNAP and 8-Br-cGMP induced endogenous Akt activation and Bad phosphorylation, resulting in the inhibition of Bad translocation to mitochondria; these effects were inhibited by KT5823 and the phosphatidylinositol 3-kinase (PI3K) inhibitors LY294002 and Wortmannin. Our data suggest that the NO/cGMP pathway suppresses 6-OHDA-induced PC 12 cell apoptosis by suppressing the mitochondrial apoptosis signal via PKG/PI3K/Akt-dependent Bad phosphorylation. - Ha, K-S., Kim, K. M., Kwon, Y.-G., Bai, S.-K., Nam, W.-D., Yoo, Y.-M., Kim, P. K. M., Chung, H.-T., Billiar, T. R., Kim, Y.-M. Nitric oxide prevents 6-hydroxydopamine-induced apoptosis in PC12 cells through cGMP-dependent PI3 kinase/Akt activation.",
author = "Ha, {Kwon Soo} and Kim, {Ki Mo} and Kwon, {Young Guen} and Bai, {Se Kyung} and Nam, {Woo Dong} and Yoo, {Young Min} and Kim, {Peter K.M.} and Chung, {Hun Taeg} and Billiar, {Timothy R.} and Kim, {Young Myeong}",
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Ha, KS, Kim, KM, Kwon, YG, Bai, SK, Nam, WD, Yoo, YM, Kim, PKM, Chung, HT, Billiar, TR & Kim, YM 2003, 'Nitric oxide prevents 6-hydroxydopamine-induced apoptosis in PC12 cells through cGMP-dependent PI3 kinase/Akt activation', FASEB Journal, vol. 17, no. 9, pp. 1036-1047. https://doi.org/10.1096/fj.02-0738com

Nitric oxide prevents 6-hydroxydopamine-induced apoptosis in PC12 cells through cGMP-dependent PI3 kinase/Akt activation. / Ha, Kwon Soo; Kim, Ki Mo; Kwon, Young Guen; Bai, Se Kyung; Nam, Woo Dong; Yoo, Young Min; Kim, Peter K.M.; Chung, Hun Taeg; Billiar, Timothy R.; Kim, Young Myeong.

In: FASEB Journal, Vol. 17, No. 9, 01.06.2003, p. 1036-1047.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Nitric oxide prevents 6-hydroxydopamine-induced apoptosis in PC12 cells through cGMP-dependent PI3 kinase/Akt activation

AU - Ha, Kwon Soo

AU - Kim, Ki Mo

AU - Kwon, Young Guen

AU - Bai, Se Kyung

AU - Nam, Woo Dong

AU - Yoo, Young Min

AU - Kim, Peter K.M.

AU - Chung, Hun Taeg

AU - Billiar, Timothy R.

AU - Kim, Young Myeong

PY - 2003/6/1

Y1 - 2003/6/1

N2 - Nitric oxide (NO) functions not only as an important signaling molecule in the brain by producing cGMP, but also regulates neuronal cell apoptosis. The mechanism by which NO regulates apoptosis is unclear. In this study, we demonstrated that NO, produced either from the NO donor S-nitroso-N-acetyl-D,L-penicillamine (SNAP) or by transfection of neuronal NO synthase, suppressed 6-hydroxydopamine (6-OHDA)-induced apoptosis in PC12 cells by inhibiting mitochondrial cytochrome c release, caspase-3 and -9 activation, and DNA fragmentation. This protection was significantly reversed by the soluble guanylyl cyclase inhibitor 1H-(1,2,4)-oxadiazole[4,3-a]quinoxalon-1-one, indicating that cGMP is a key mediator in NO-mediated anti-apoptosis. Moreover, the membrane-permeable cGMP analog 8-Br-cGMP inhibited 6-OHDA-induced apoptosis. These anti-apoptotic effects of SNAP and 8-Br-cGMP were suppressed by cGMP-dependent protein kinase G (PKG) inhibitor KT5823, indicating that PKG is a downstream signal mediator in the suppression of apoptosis by NO and cGMP. Both SNAP and 8-Br-cGMP induced endogenous Akt activation and Bad phosphorylation, resulting in the inhibition of Bad translocation to mitochondria; these effects were inhibited by KT5823 and the phosphatidylinositol 3-kinase (PI3K) inhibitors LY294002 and Wortmannin. Our data suggest that the NO/cGMP pathway suppresses 6-OHDA-induced PC 12 cell apoptosis by suppressing the mitochondrial apoptosis signal via PKG/PI3K/Akt-dependent Bad phosphorylation. - Ha, K-S., Kim, K. M., Kwon, Y.-G., Bai, S.-K., Nam, W.-D., Yoo, Y.-M., Kim, P. K. M., Chung, H.-T., Billiar, T. R., Kim, Y.-M. Nitric oxide prevents 6-hydroxydopamine-induced apoptosis in PC12 cells through cGMP-dependent PI3 kinase/Akt activation.

AB - Nitric oxide (NO) functions not only as an important signaling molecule in the brain by producing cGMP, but also regulates neuronal cell apoptosis. The mechanism by which NO regulates apoptosis is unclear. In this study, we demonstrated that NO, produced either from the NO donor S-nitroso-N-acetyl-D,L-penicillamine (SNAP) or by transfection of neuronal NO synthase, suppressed 6-hydroxydopamine (6-OHDA)-induced apoptosis in PC12 cells by inhibiting mitochondrial cytochrome c release, caspase-3 and -9 activation, and DNA fragmentation. This protection was significantly reversed by the soluble guanylyl cyclase inhibitor 1H-(1,2,4)-oxadiazole[4,3-a]quinoxalon-1-one, indicating that cGMP is a key mediator in NO-mediated anti-apoptosis. Moreover, the membrane-permeable cGMP analog 8-Br-cGMP inhibited 6-OHDA-induced apoptosis. These anti-apoptotic effects of SNAP and 8-Br-cGMP were suppressed by cGMP-dependent protein kinase G (PKG) inhibitor KT5823, indicating that PKG is a downstream signal mediator in the suppression of apoptosis by NO and cGMP. Both SNAP and 8-Br-cGMP induced endogenous Akt activation and Bad phosphorylation, resulting in the inhibition of Bad translocation to mitochondria; these effects were inhibited by KT5823 and the phosphatidylinositol 3-kinase (PI3K) inhibitors LY294002 and Wortmannin. Our data suggest that the NO/cGMP pathway suppresses 6-OHDA-induced PC 12 cell apoptosis by suppressing the mitochondrial apoptosis signal via PKG/PI3K/Akt-dependent Bad phosphorylation. - Ha, K-S., Kim, K. M., Kwon, Y.-G., Bai, S.-K., Nam, W.-D., Yoo, Y.-M., Kim, P. K. M., Chung, H.-T., Billiar, T. R., Kim, Y.-M. Nitric oxide prevents 6-hydroxydopamine-induced apoptosis in PC12 cells through cGMP-dependent PI3 kinase/Akt activation.

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