Involvement of sphingosine-1-Phosphate/RhoA/Rho-Kinase signaling pathway in corporal fibrosis following cavernous nerve injury in male rats

Min Chul Cho, Kwanjin Park, Ji Sun Chai, Sun Hee Lee, Soowoong Kim, Jae Seung Paick

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

Introduction. Postprostatectomy erectile dysfunction (ED) is thought to be due primarily to injury to cavernous nerve (CN) during surgery. The molecular mechanisms leading to ED after CN injury are poorly understood. Aim. We determined whether transforming growth factor-β1 (TGF-β1), sphingosine-1-phosphate (S1P) and RhoA/Rho-kinase (ROCK) signaling pathways were involved in corporal fibrosis after bilateral CN injury in rats. Methods. Forty-eight 10-week-old male Sprague-Dawley rats were equally divided into the following four groups: normal control group (C); sham surgery group (S); bilateral CN crush injury group (I); and bilateral CN transection group (T). Within each of the four groups, two subgroups were analyzed as a function of time (1 and 8 weeks postoperatively). Main Outcome Measures. Penile tissue was processed for immunoblot (RhoA, ROCK1, phospho-myosin phosphatase target subunit [MYPT1]), reverse transcription-polymerase chain reaction (RT-PCR) (TGF-β1, sphingosine kinase type 1 [SphK1], and S1P2), immunohistochemistry (alpha smooth muscle actin [α-SMA]), and Masson's trichrome staining. Results. At 1 and 8 weeks postoperatively, the I and T groups had a significantly decreased smooth muscle cell/collagen ratio, the expression of α-SMA and phospho-MYPT1 compared to the C group. Densitometry revealed a significantly higher expression of RhoA and ROCK1 in the T group compared to the C group at 1 and 8 weeks postoperatively. For the I group, the expression of RhoA significantly increased starting from 1 week postoperatively, but the expression of ROCK1 significantly increased as late as 8 weeks following injury. The expression of TGF-β1 and S1P2 mRNA in the I or T group remained significantly increased up to 8 weeks compared to the C group, despite significant reduction at 8 weeks compared to 1 week postoperatively. The expression of SphK1 mRNA in the I and T groups was significantly increased at 1 week but not 8 weeks postoperatively. Conclusions. Our data suggest that S1P and RhoA/ROCK1 signaling may be involved in corporal fibrosis associated with loss of smooth muscle through coordination with TGF-β1 after CN injury.

Original languageEnglish
Pages (from-to)712-721
Number of pages10
JournalJournal of Sexual Medicine
Volume8
Issue number3
DOIs
StatePublished - 1 Jan 2011

Fingerprint

rho-Associated Kinases
Transforming Growth Factors
Fibrosis
Wounds and Injuries
Erectile Dysfunction
Smooth Muscle
Myosin-Light-Chain Phosphatase
Nerve Crush
Messenger RNA
Densitometry
Reverse Transcription
Smooth Muscle Myocytes
Sprague Dawley Rats
Actins
Collagen
Immunohistochemistry
Outcome Assessment (Health Care)
sphingosine 1-phosphate
Staining and Labeling
Polymerase Chain Reaction

Keywords

  • Fibrosis
  • Penis
  • Postprostatectomy Erectile Dysfunction
  • Rho Kinase
  • RhoA
  • S1P Receptor
  • Sphingosine Kinase
  • TGF-Beta

Cite this

@article{2ce1b758f05f4ad78e41ac53a9aacb81,
title = "Involvement of sphingosine-1-Phosphate/RhoA/Rho-Kinase signaling pathway in corporal fibrosis following cavernous nerve injury in male rats",
abstract = "Introduction. Postprostatectomy erectile dysfunction (ED) is thought to be due primarily to injury to cavernous nerve (CN) during surgery. The molecular mechanisms leading to ED after CN injury are poorly understood. Aim. We determined whether transforming growth factor-β1 (TGF-β1), sphingosine-1-phosphate (S1P) and RhoA/Rho-kinase (ROCK) signaling pathways were involved in corporal fibrosis after bilateral CN injury in rats. Methods. Forty-eight 10-week-old male Sprague-Dawley rats were equally divided into the following four groups: normal control group (C); sham surgery group (S); bilateral CN crush injury group (I); and bilateral CN transection group (T). Within each of the four groups, two subgroups were analyzed as a function of time (1 and 8 weeks postoperatively). Main Outcome Measures. Penile tissue was processed for immunoblot (RhoA, ROCK1, phospho-myosin phosphatase target subunit [MYPT1]), reverse transcription-polymerase chain reaction (RT-PCR) (TGF-β1, sphingosine kinase type 1 [SphK1], and S1P2), immunohistochemistry (alpha smooth muscle actin [α-SMA]), and Masson's trichrome staining. Results. At 1 and 8 weeks postoperatively, the I and T groups had a significantly decreased smooth muscle cell/collagen ratio, the expression of α-SMA and phospho-MYPT1 compared to the C group. Densitometry revealed a significantly higher expression of RhoA and ROCK1 in the T group compared to the C group at 1 and 8 weeks postoperatively. For the I group, the expression of RhoA significantly increased starting from 1 week postoperatively, but the expression of ROCK1 significantly increased as late as 8 weeks following injury. The expression of TGF-β1 and S1P2 mRNA in the I or T group remained significantly increased up to 8 weeks compared to the C group, despite significant reduction at 8 weeks compared to 1 week postoperatively. The expression of SphK1 mRNA in the I and T groups was significantly increased at 1 week but not 8 weeks postoperatively. Conclusions. Our data suggest that S1P and RhoA/ROCK1 signaling may be involved in corporal fibrosis associated with loss of smooth muscle through coordination with TGF-β1 after CN injury.",
keywords = "Fibrosis, Penis, Postprostatectomy Erectile Dysfunction, Rho Kinase, RhoA, S1P Receptor, Sphingosine Kinase, TGF-Beta",
author = "Cho, {Min Chul} and Kwanjin Park and Chai, {Ji Sun} and Lee, {Sun Hee} and Soowoong Kim and Paick, {Jae Seung}",
year = "2011",
month = "1",
day = "1",
doi = "10.1111/j.1743-6109.2010.02147.x",
language = "English",
volume = "8",
pages = "712--721",
journal = "Journal of Sexual Medicine",
issn = "1743-6095",
publisher = "Elsevier Inc.",
number = "3",

}

Involvement of sphingosine-1-Phosphate/RhoA/Rho-Kinase signaling pathway in corporal fibrosis following cavernous nerve injury in male rats. / Cho, Min Chul; Park, Kwanjin; Chai, Ji Sun; Lee, Sun Hee; Kim, Soowoong; Paick, Jae Seung.

In: Journal of Sexual Medicine, Vol. 8, No. 3, 01.01.2011, p. 712-721.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Involvement of sphingosine-1-Phosphate/RhoA/Rho-Kinase signaling pathway in corporal fibrosis following cavernous nerve injury in male rats

AU - Cho, Min Chul

AU - Park, Kwanjin

AU - Chai, Ji Sun

AU - Lee, Sun Hee

AU - Kim, Soowoong

AU - Paick, Jae Seung

PY - 2011/1/1

Y1 - 2011/1/1

N2 - Introduction. Postprostatectomy erectile dysfunction (ED) is thought to be due primarily to injury to cavernous nerve (CN) during surgery. The molecular mechanisms leading to ED after CN injury are poorly understood. Aim. We determined whether transforming growth factor-β1 (TGF-β1), sphingosine-1-phosphate (S1P) and RhoA/Rho-kinase (ROCK) signaling pathways were involved in corporal fibrosis after bilateral CN injury in rats. Methods. Forty-eight 10-week-old male Sprague-Dawley rats were equally divided into the following four groups: normal control group (C); sham surgery group (S); bilateral CN crush injury group (I); and bilateral CN transection group (T). Within each of the four groups, two subgroups were analyzed as a function of time (1 and 8 weeks postoperatively). Main Outcome Measures. Penile tissue was processed for immunoblot (RhoA, ROCK1, phospho-myosin phosphatase target subunit [MYPT1]), reverse transcription-polymerase chain reaction (RT-PCR) (TGF-β1, sphingosine kinase type 1 [SphK1], and S1P2), immunohistochemistry (alpha smooth muscle actin [α-SMA]), and Masson's trichrome staining. Results. At 1 and 8 weeks postoperatively, the I and T groups had a significantly decreased smooth muscle cell/collagen ratio, the expression of α-SMA and phospho-MYPT1 compared to the C group. Densitometry revealed a significantly higher expression of RhoA and ROCK1 in the T group compared to the C group at 1 and 8 weeks postoperatively. For the I group, the expression of RhoA significantly increased starting from 1 week postoperatively, but the expression of ROCK1 significantly increased as late as 8 weeks following injury. The expression of TGF-β1 and S1P2 mRNA in the I or T group remained significantly increased up to 8 weeks compared to the C group, despite significant reduction at 8 weeks compared to 1 week postoperatively. The expression of SphK1 mRNA in the I and T groups was significantly increased at 1 week but not 8 weeks postoperatively. Conclusions. Our data suggest that S1P and RhoA/ROCK1 signaling may be involved in corporal fibrosis associated with loss of smooth muscle through coordination with TGF-β1 after CN injury.

AB - Introduction. Postprostatectomy erectile dysfunction (ED) is thought to be due primarily to injury to cavernous nerve (CN) during surgery. The molecular mechanisms leading to ED after CN injury are poorly understood. Aim. We determined whether transforming growth factor-β1 (TGF-β1), sphingosine-1-phosphate (S1P) and RhoA/Rho-kinase (ROCK) signaling pathways were involved in corporal fibrosis after bilateral CN injury in rats. Methods. Forty-eight 10-week-old male Sprague-Dawley rats were equally divided into the following four groups: normal control group (C); sham surgery group (S); bilateral CN crush injury group (I); and bilateral CN transection group (T). Within each of the four groups, two subgroups were analyzed as a function of time (1 and 8 weeks postoperatively). Main Outcome Measures. Penile tissue was processed for immunoblot (RhoA, ROCK1, phospho-myosin phosphatase target subunit [MYPT1]), reverse transcription-polymerase chain reaction (RT-PCR) (TGF-β1, sphingosine kinase type 1 [SphK1], and S1P2), immunohistochemistry (alpha smooth muscle actin [α-SMA]), and Masson's trichrome staining. Results. At 1 and 8 weeks postoperatively, the I and T groups had a significantly decreased smooth muscle cell/collagen ratio, the expression of α-SMA and phospho-MYPT1 compared to the C group. Densitometry revealed a significantly higher expression of RhoA and ROCK1 in the T group compared to the C group at 1 and 8 weeks postoperatively. For the I group, the expression of RhoA significantly increased starting from 1 week postoperatively, but the expression of ROCK1 significantly increased as late as 8 weeks following injury. The expression of TGF-β1 and S1P2 mRNA in the I or T group remained significantly increased up to 8 weeks compared to the C group, despite significant reduction at 8 weeks compared to 1 week postoperatively. The expression of SphK1 mRNA in the I and T groups was significantly increased at 1 week but not 8 weeks postoperatively. Conclusions. Our data suggest that S1P and RhoA/ROCK1 signaling may be involved in corporal fibrosis associated with loss of smooth muscle through coordination with TGF-β1 after CN injury.

KW - Fibrosis

KW - Penis

KW - Postprostatectomy Erectile Dysfunction

KW - Rho Kinase

KW - RhoA

KW - S1P Receptor

KW - Sphingosine Kinase

KW - TGF-Beta

UR - http://www.scopus.com/inward/record.url?scp=79951977947&partnerID=8YFLogxK

U2 - 10.1111/j.1743-6109.2010.02147.x

DO - 10.1111/j.1743-6109.2010.02147.x

M3 - Article

C2 - 21143420

AN - SCOPUS:79951977947

VL - 8

SP - 712

EP - 721

JO - Journal of Sexual Medicine

JF - Journal of Sexual Medicine

SN - 1743-6095

IS - 3

ER -