Endothelin-1 Upregulates Activin Receptor-Like Kinase-1 Expression Via G/RhoA/Sp-1/Rho Kinase Pathways in Human Pulmonary Arterial Endothelial Cells

Overview

Journal Front Cardiovasc Med

Specialty Cardiology & Vascular Diseases

Date 2021 Mar 12

PMID 33708809

Citations 1

Authors

Koichi Sugimoto

Tetsuro Yokokawa

Tomofumi Misaka

Takashi Kaneshiro

Shinya Yamada

Akiomi Yoshihisa

Kazuhiko Nakazato

Yasuchika Takeishi

Affiliations

Soon will be listed here.

Abstract

Pulmonary arterial hypertension (PAH) is characterized by pulmonary vasoconstriction and organic stenosis. It has been demonstrated that endothelin-1 (ET-1) induces pulmonary vasoconstriction through the activation of RhoA. In addition, a gene mutation of activin receptor-like kinase (ACVRL)-1 is recognized in PAH patients. However, little is known about the association between ET-1 and ACVRL-1. In the present study, we aimed to investigate the effect of ET-1 on ACVRL-1 expression and delineate the involvement of the G/RhoA/Rho kinase pathway. ET-1 was added to culture medium of human pulmonary arterial endothelial cells (PAECs). Pre-treatment with pertussis toxin (PTX) or exoenzyme C3 transferase (C3T) was performed for inhibition of G or RhoA, respectively. Rho kinase was inhibited by Y27632. Mithramycin A was used for inhibition of Sp-1, which is a transcriptional factor of ACVRL-1. The active form of RhoA (GTP-RhoA) was assessed by pull-down assay. ACVRL-1 expression was increased by ET-1 in the PAECs. Pull-down assay revealed that ET-1 induced GTP-loading of RhoA, which was suppressed by pre-treatment with PTX or C3T. Further, PTX, C3T, and Y27632 suppressed the ET-1-induced ACVRL-1 expression. ET-1 increased the activity of the ACVRL-1 promoter and stabilized the ACVRL-1 mRNA. Sp-1 peaked 15 min after adding ET-1 to the PAECs. PTX and C3T prevented the increase of Sp-1 induced by ET-1. Inhibition of Sp-1 by mithramycin A suppressed ET-1-induced ACVRL-1 upregulation. The present study demonstrated that ET-1 increases ACVRL-1 expression in human PAECs the G/RhoA/Rho kinase pathway with the involvement of Sp-1.

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