Regulation of endothelial production of C-type natriuretic peptide in coculture with vascular smooth muscle cells

Role of the vascular natriuretic peptide system in vascular growth inhibition

Yasato Komatsu, Hiroshi Itoh, Shin Ichi Suga, Yoshihiro Ogawa, Norio Hama, Ichiro Kishimoto, Osamu Nakagawa, Toshio Igaki, Kentaro Doi, Takaaki Yoshimasa, Kazuwa Nakao

Research output: Contribution to journalArticle

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Abstract

Recently, we have demonstrated that C-type natriuretic peptide (CNP) is produced in vascular endothelial cells (ECs). In the present study, we investigated the interaction of ECs and vascular smooth muscle cells (SMCs) for endothelial production of CNP and its action on vascular growth, using the EC/SMC coculture system. The concentration of CNP-like immunoreactivity in the medium was increased 60-fold within 48 hours in the EC/SMC coculture with direct contact compared with that in EC alone. Northern blot analysis revealed the augmented expression of CNP mRNA in the EC/SMC coculture. The accumulation of intracellular cGMP in the coculture was concomitantly increased, and this response was blocked by anti-CNP monoclonal antibody and HS-142-1, a nonpeptide atrial natriuretic peptide receptor antagonist. The concentration of biologically active transforming growth factor-β (TGF-β) in the culture medium of the coculture with direct contact of ECs and SMCs was elevated to the level to stimulate endothelial production of CNP. Actually, the neutralizing antibody against TGF-β abrogated the cGMP accumulation in the coculture. These results show that endothelial production of CNP in the EC/SMC coculture is at least in part regulated by TGF-β. Furthermore, the conditioned medium from ECs stimulated by TGF-β was demonstrated to have a growth-inhibitory effect on SMCs, which was abolished by anti-CNP monoclonal antibody and HS-142-1. The treatment with anti-CNP monoclonal antibody and HS-142-1 also significantly increased the cell number of the EC/SMC coculture. The present study reveals the pathophysiological significance of endothelial CNP as a paracrine/autocrine vascular regulator for vascular growth in the interaction of ECs and SMCs.

Original languageEnglish
Pages (from-to)606-614
Number of pages9
JournalCirculation research
Volume78
Issue number4
DOIs
Publication statusPublished - Jan 1 1996
Externally publishedYes

Fingerprint

C-Type Natriuretic Peptide
Natriuretic Peptides
Coculture Techniques
Vascular Smooth Muscle
Smooth Muscle Myocytes
Blood Vessels
Endothelial Cells
Growth
Transforming Growth Factors
Monoclonal Antibodies
Atrial Natriuretic Factor Receptors
Conditioned Culture Medium
Neutralizing Antibodies
Northern Blotting
Culture Media

All Science Journal Classification (ASJC) codes

  • Physiology
  • Cardiology and Cardiovascular Medicine

Cite this

Regulation of endothelial production of C-type natriuretic peptide in coculture with vascular smooth muscle cells : Role of the vascular natriuretic peptide system in vascular growth inhibition. / Komatsu, Yasato; Itoh, Hiroshi; Suga, Shin Ichi; Ogawa, Yoshihiro; Hama, Norio; Kishimoto, Ichiro; Nakagawa, Osamu; Igaki, Toshio; Doi, Kentaro; Yoshimasa, Takaaki; Nakao, Kazuwa.

In: Circulation research, Vol. 78, No. 4, 01.01.1996, p. 606-614.

Research output: Contribution to journalArticle

Komatsu, Yasato ; Itoh, Hiroshi ; Suga, Shin Ichi ; Ogawa, Yoshihiro ; Hama, Norio ; Kishimoto, Ichiro ; Nakagawa, Osamu ; Igaki, Toshio ; Doi, Kentaro ; Yoshimasa, Takaaki ; Nakao, Kazuwa. / Regulation of endothelial production of C-type natriuretic peptide in coculture with vascular smooth muscle cells : Role of the vascular natriuretic peptide system in vascular growth inhibition. In: Circulation research. 1996 ; Vol. 78, No. 4. pp. 606-614.
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abstract = "Recently, we have demonstrated that C-type natriuretic peptide (CNP) is produced in vascular endothelial cells (ECs). In the present study, we investigated the interaction of ECs and vascular smooth muscle cells (SMCs) for endothelial production of CNP and its action on vascular growth, using the EC/SMC coculture system. The concentration of CNP-like immunoreactivity in the medium was increased 60-fold within 48 hours in the EC/SMC coculture with direct contact compared with that in EC alone. Northern blot analysis revealed the augmented expression of CNP mRNA in the EC/SMC coculture. The accumulation of intracellular cGMP in the coculture was concomitantly increased, and this response was blocked by anti-CNP monoclonal antibody and HS-142-1, a nonpeptide atrial natriuretic peptide receptor antagonist. The concentration of biologically active transforming growth factor-β (TGF-β) in the culture medium of the coculture with direct contact of ECs and SMCs was elevated to the level to stimulate endothelial production of CNP. Actually, the neutralizing antibody against TGF-β abrogated the cGMP accumulation in the coculture. These results show that endothelial production of CNP in the EC/SMC coculture is at least in part regulated by TGF-β. Furthermore, the conditioned medium from ECs stimulated by TGF-β was demonstrated to have a growth-inhibitory effect on SMCs, which was abolished by anti-CNP monoclonal antibody and HS-142-1. The treatment with anti-CNP monoclonal antibody and HS-142-1 also significantly increased the cell number of the EC/SMC coculture. The present study reveals the pathophysiological significance of endothelial CNP as a paracrine/autocrine vascular regulator for vascular growth in the interaction of ECs and SMCs.",
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AU - Komatsu, Yasato

AU - Itoh, Hiroshi

AU - Suga, Shin Ichi

AU - Ogawa, Yoshihiro

AU - Hama, Norio

AU - Kishimoto, Ichiro

AU - Nakagawa, Osamu

AU - Igaki, Toshio

AU - Doi, Kentaro

AU - Yoshimasa, Takaaki

AU - Nakao, Kazuwa

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