Corpus cavernosal smooth muscle relaxation effect of a novel AMPK activator, beta-lapachone

Jae Hyun Bae, Jin Wook Kim, Gi Ryang Kweon, Myoung Gyu Park, Kyeong Hoon Jeong, Je-Jong Kim, Du Geon Moon

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Introduction. Adenosine monophosphate-activated protein kinase (AMPK) activation is suggested to relax smooth muscle by endothelial nitric oxide synthase (eNOS) phosphorylation. Aim. To assess the mechanism and effect of a novel AMPK activator, beta-lapachone, upon cavernosal smooth muscle relaxation and the therapeutic potential for erectile dysfunction. Methods. Human umbilical vein endothelial cells (HUVECs) were treated with beta-lapachone. The lysates were blotted with specific antibodies for phosphorylated AMPK (p-AMPK) or phosphorylated eNOS (p-eNOS). The membranes were re-blotted for total AMP total eNOS, or beta-actin. The eNOS activity was measured by the conversion of L-14C-arginine to L-14C-citrulline in HUVECs lysates. In a separated experiment, cavernosal strips from New Zealand white rabbits were harvested for organ bath study and the relaxation effect of beta-lapachone on phenylephrine-induced contracted strips was evaluated and compared with sodium nitroprusside, zaprinast, metformin, and aminoimidazole carboxamide ribonucleotide (AICAR). Methylene blue and L-NAME were used to assess the inhibition of cyclic guanosine monophosphate/nitric oxide pathway. Zinc-protoporphyrin-IX (ZnPP) was also used to investigate the contribution of mevalonate pathway. Main Outcome Measures. The expression of p-AMPK, p-eNOS, AMPK and eNOS induced by beta-lapachone in HUVECs study and the percent relaxation of cavernosal tissue in organ bath study. Results. Beta-lapachone clearly induced AMPK phosphorylation and, as a consequence, eNOS phosphorylation in HUVECs. Beta-lapachone-induced upregulation of eNOS activity was also observed in HUVECs and steadily increased up to 1 hour. In organ bath study, beta-lapachone significantly relaxed the phenylephrine pretreated strips in a dose-dependent manner. This relaxation effect was not totally blocked by methylene blue or L-NAME. After removing endothelium, the relaxation was totally blocked by ZnPP. Conclusions. A novel AMPK activator, beta-lapachone has a strong relaxation effect on precontracted cavernosal smooth muscle strips in the rabbit. And phosphorylation of AMPK and eNOS strongly related to the action of beta-lapachone. Mevalonate pathway also might be considered as a suggestive mechanism.

Original languageEnglish
Pages (from-to)2205-2214
Number of pages10
JournalJournal of Sexual Medicine
Volume8
Issue number8
DOIs
Publication statusPublished - 2011 Jan 1

Fingerprint

Muscle Relaxation
Adenosine Monophosphate
Nitric Oxide Synthase Type III
Protein Kinases
Smooth Muscle
Human Umbilical Vein Endothelial Cells
Baths
Phosphorylation
Mevalonic Acid
Methylene Blue
NG-Nitroarginine Methyl Ester
Phenylephrine
Rabbits
beta-lapachone
Citrulline
AMP-Activated Protein Kinases
Metformin
Cyclic GMP
Nitroprusside
Erectile Dysfunction

Keywords

  • AMPK Activator
  • Beta-Lapachone
  • Corpus Cavernosum
  • ENOS
  • Penile Erection
  • Smooth Muscle Relaxation

ASJC Scopus subject areas

  • Reproductive Medicine
  • Obstetrics and Gynaecology
  • Urology

Cite this

Corpus cavernosal smooth muscle relaxation effect of a novel AMPK activator, beta-lapachone. / Bae, Jae Hyun; Kim, Jin Wook; Kweon, Gi Ryang; Park, Myoung Gyu; Jeong, Kyeong Hoon; Kim, Je-Jong; Moon, Du Geon.

In: Journal of Sexual Medicine, Vol. 8, No. 8, 01.01.2011, p. 2205-2214.

Research output: Contribution to journalArticle

Bae, Jae Hyun ; Kim, Jin Wook ; Kweon, Gi Ryang ; Park, Myoung Gyu ; Jeong, Kyeong Hoon ; Kim, Je-Jong ; Moon, Du Geon. / Corpus cavernosal smooth muscle relaxation effect of a novel AMPK activator, beta-lapachone. In: Journal of Sexual Medicine. 2011 ; Vol. 8, No. 8. pp. 2205-2214.
@article{84d2c3d320da4c7b96e1af191f69af14,
title = "Corpus cavernosal smooth muscle relaxation effect of a novel AMPK activator, beta-lapachone",
abstract = "Introduction. Adenosine monophosphate-activated protein kinase (AMPK) activation is suggested to relax smooth muscle by endothelial nitric oxide synthase (eNOS) phosphorylation. Aim. To assess the mechanism and effect of a novel AMPK activator, beta-lapachone, upon cavernosal smooth muscle relaxation and the therapeutic potential for erectile dysfunction. Methods. Human umbilical vein endothelial cells (HUVECs) were treated with beta-lapachone. The lysates were blotted with specific antibodies for phosphorylated AMPK (p-AMPK) or phosphorylated eNOS (p-eNOS). The membranes were re-blotted for total AMP total eNOS, or beta-actin. The eNOS activity was measured by the conversion of L-14C-arginine to L-14C-citrulline in HUVECs lysates. In a separated experiment, cavernosal strips from New Zealand white rabbits were harvested for organ bath study and the relaxation effect of beta-lapachone on phenylephrine-induced contracted strips was evaluated and compared with sodium nitroprusside, zaprinast, metformin, and aminoimidazole carboxamide ribonucleotide (AICAR). Methylene blue and L-NAME were used to assess the inhibition of cyclic guanosine monophosphate/nitric oxide pathway. Zinc-protoporphyrin-IX (ZnPP) was also used to investigate the contribution of mevalonate pathway. Main Outcome Measures. The expression of p-AMPK, p-eNOS, AMPK and eNOS induced by beta-lapachone in HUVECs study and the percent relaxation of cavernosal tissue in organ bath study. Results. Beta-lapachone clearly induced AMPK phosphorylation and, as a consequence, eNOS phosphorylation in HUVECs. Beta-lapachone-induced upregulation of eNOS activity was also observed in HUVECs and steadily increased up to 1 hour. In organ bath study, beta-lapachone significantly relaxed the phenylephrine pretreated strips in a dose-dependent manner. This relaxation effect was not totally blocked by methylene blue or L-NAME. After removing endothelium, the relaxation was totally blocked by ZnPP. Conclusions. A novel AMPK activator, beta-lapachone has a strong relaxation effect on precontracted cavernosal smooth muscle strips in the rabbit. And phosphorylation of AMPK and eNOS strongly related to the action of beta-lapachone. Mevalonate pathway also might be considered as a suggestive mechanism.",
keywords = "AMPK Activator, Beta-Lapachone, Corpus Cavernosum, ENOS, Penile Erection, Smooth Muscle Relaxation",
author = "Bae, {Jae Hyun} and Kim, {Jin Wook} and Kweon, {Gi Ryang} and Park, {Myoung Gyu} and Jeong, {Kyeong Hoon} and Je-Jong Kim and Moon, {Du Geon}",
year = "2011",
month = "1",
day = "1",
doi = "10.1111/j.1743-6109.2010.01809.x",
language = "English",
volume = "8",
pages = "2205--2214",
journal = "Journal of Sexual Medicine",
issn = "1743-6095",
publisher = "Wiley-Blackwell",
number = "8",

}

TY - JOUR

T1 - Corpus cavernosal smooth muscle relaxation effect of a novel AMPK activator, beta-lapachone

AU - Bae, Jae Hyun

AU - Kim, Jin Wook

AU - Kweon, Gi Ryang

AU - Park, Myoung Gyu

AU - Jeong, Kyeong Hoon

AU - Kim, Je-Jong

AU - Moon, Du Geon

PY - 2011/1/1

Y1 - 2011/1/1

N2 - Introduction. Adenosine monophosphate-activated protein kinase (AMPK) activation is suggested to relax smooth muscle by endothelial nitric oxide synthase (eNOS) phosphorylation. Aim. To assess the mechanism and effect of a novel AMPK activator, beta-lapachone, upon cavernosal smooth muscle relaxation and the therapeutic potential for erectile dysfunction. Methods. Human umbilical vein endothelial cells (HUVECs) were treated with beta-lapachone. The lysates were blotted with specific antibodies for phosphorylated AMPK (p-AMPK) or phosphorylated eNOS (p-eNOS). The membranes were re-blotted for total AMP total eNOS, or beta-actin. The eNOS activity was measured by the conversion of L-14C-arginine to L-14C-citrulline in HUVECs lysates. In a separated experiment, cavernosal strips from New Zealand white rabbits were harvested for organ bath study and the relaxation effect of beta-lapachone on phenylephrine-induced contracted strips was evaluated and compared with sodium nitroprusside, zaprinast, metformin, and aminoimidazole carboxamide ribonucleotide (AICAR). Methylene blue and L-NAME were used to assess the inhibition of cyclic guanosine monophosphate/nitric oxide pathway. Zinc-protoporphyrin-IX (ZnPP) was also used to investigate the contribution of mevalonate pathway. Main Outcome Measures. The expression of p-AMPK, p-eNOS, AMPK and eNOS induced by beta-lapachone in HUVECs study and the percent relaxation of cavernosal tissue in organ bath study. Results. Beta-lapachone clearly induced AMPK phosphorylation and, as a consequence, eNOS phosphorylation in HUVECs. Beta-lapachone-induced upregulation of eNOS activity was also observed in HUVECs and steadily increased up to 1 hour. In organ bath study, beta-lapachone significantly relaxed the phenylephrine pretreated strips in a dose-dependent manner. This relaxation effect was not totally blocked by methylene blue or L-NAME. After removing endothelium, the relaxation was totally blocked by ZnPP. Conclusions. A novel AMPK activator, beta-lapachone has a strong relaxation effect on precontracted cavernosal smooth muscle strips in the rabbit. And phosphorylation of AMPK and eNOS strongly related to the action of beta-lapachone. Mevalonate pathway also might be considered as a suggestive mechanism.

AB - Introduction. Adenosine monophosphate-activated protein kinase (AMPK) activation is suggested to relax smooth muscle by endothelial nitric oxide synthase (eNOS) phosphorylation. Aim. To assess the mechanism and effect of a novel AMPK activator, beta-lapachone, upon cavernosal smooth muscle relaxation and the therapeutic potential for erectile dysfunction. Methods. Human umbilical vein endothelial cells (HUVECs) were treated with beta-lapachone. The lysates were blotted with specific antibodies for phosphorylated AMPK (p-AMPK) or phosphorylated eNOS (p-eNOS). The membranes were re-blotted for total AMP total eNOS, or beta-actin. The eNOS activity was measured by the conversion of L-14C-arginine to L-14C-citrulline in HUVECs lysates. In a separated experiment, cavernosal strips from New Zealand white rabbits were harvested for organ bath study and the relaxation effect of beta-lapachone on phenylephrine-induced contracted strips was evaluated and compared with sodium nitroprusside, zaprinast, metformin, and aminoimidazole carboxamide ribonucleotide (AICAR). Methylene blue and L-NAME were used to assess the inhibition of cyclic guanosine monophosphate/nitric oxide pathway. Zinc-protoporphyrin-IX (ZnPP) was also used to investigate the contribution of mevalonate pathway. Main Outcome Measures. The expression of p-AMPK, p-eNOS, AMPK and eNOS induced by beta-lapachone in HUVECs study and the percent relaxation of cavernosal tissue in organ bath study. Results. Beta-lapachone clearly induced AMPK phosphorylation and, as a consequence, eNOS phosphorylation in HUVECs. Beta-lapachone-induced upregulation of eNOS activity was also observed in HUVECs and steadily increased up to 1 hour. In organ bath study, beta-lapachone significantly relaxed the phenylephrine pretreated strips in a dose-dependent manner. This relaxation effect was not totally blocked by methylene blue or L-NAME. After removing endothelium, the relaxation was totally blocked by ZnPP. Conclusions. A novel AMPK activator, beta-lapachone has a strong relaxation effect on precontracted cavernosal smooth muscle strips in the rabbit. And phosphorylation of AMPK and eNOS strongly related to the action of beta-lapachone. Mevalonate pathway also might be considered as a suggestive mechanism.

KW - AMPK Activator

KW - Beta-Lapachone

KW - Corpus Cavernosum

KW - ENOS

KW - Penile Erection

KW - Smooth Muscle Relaxation

UR - http://www.scopus.com/inward/record.url?scp=79960829688&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=79960829688&partnerID=8YFLogxK

U2 - 10.1111/j.1743-6109.2010.01809.x

DO - 10.1111/j.1743-6109.2010.01809.x

M3 - Article

VL - 8

SP - 2205

EP - 2214

JO - Journal of Sexual Medicine

JF - Journal of Sexual Medicine

SN - 1743-6095

IS - 8

ER -