CCN3 secreted by prostaglandin E2 inhibits intimal cushion formation in the rat ductus arteriosus

Journal Article · Sat Sep 15 04:00:00 EDT 2018 · Biochemical and Biophysical Research Communications

DOI:https://doi.org/10.1016/J.BBRC.2018.08.138· OSTI ID:23103572

Iwai, Kenji ^[1]; Nagasawa, Kazumichi ^[2]; Akaike, Toru ^[3]; Oshima, Toshio ^[1]; Kato, Takashi ^[2]; Minamisawa, Susumu ^[1]

---

1. Graduate School of Life Science and Medical Bioscience, Waseda University, Tokyo (Japan)
2. Graduate School of Science and Engineering, Bioscience and Biomedical Engineering, Waseda Univeristy, Tokyo (Japan)
3. Department of Cell Physiology, The Jikei University School of Medicine, Tokyo (Japan)

Highlights: • CCN3 is secreted from smooth muscle cells of the ductus arteriosus by prostaglandin E{sub 2}. • Nano-flow LC-MS/MS analysis is a powerful tool for detecting a novel secreted protein. • CCN3 inhibits intimal cushion formation in the ductus arteriosus. The ductus arteriosus (DA), an essential fetal shunt between the pulmonary trunk and the descending aorta, changes its structure during development. Our previous studies have demonstrated that prostaglandin E{sub 2} (PGE{sub 2})-EP4 signaling promotes intimal cushion formation (ICF) by activating the migration of DA smooth muscle cells via the secretion of hyaluronan. We hypothesized that, in addition to hyaluronan, PGE{sub 2} may secrete other proteins that also regulate vascular remodeling in the DA. In order to detect PGE{sub 2} stimulation-secreted proteins, we found that CCN3 protein was increased in the culture supernatant in the presence of PGE{sub 2} in a dose-dependent manner by nano-flow liquid chromatography coupled with tandem mass spectrometry analysis and enzyme-linked immunosorbent assay. Quantitative RT-PCR analysis revealed that PGE{sub 2} stimulation tended to increase the expression levels of CCN3 mRNA in DA smooth muscle cells. Immunohistochemical analysis revealed that CCN3 was highly localized in the entire smooth muscle layers and the endothelium of the DA. Furthermore, exogenous CCN3 inhibited PGE{sub 2}-induced ICF in the ex vivo DA tissues. These results suggest that CCN3 is a secreted protein of the DA smooth muscle cells induced by PGE{sub 2} to suppress ICF of the DA. The present study indicates that CCN3 could be a novel negative regulator of ICF in the DA to fine-tune the PGE{sub 2}-mediated DA remodeling.

• OSTI ID: 23103572
• Journal Information: Biochemical and Biophysical Research Communications, Journal Name: Biochemical and Biophysical Research Communications Journal Issue: 4 Vol. 503; ISSN BBRCA9; ISSN 0006-291X
• Country of Publication: United States
• Language: English

Related Subjects

60 APPLIED LIFE SCIENCES
AORTA
ENDOTHELIUM
ENZYMES
MESSENGER-RNA
MUSCLES
RATS