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Mechanical strain increases endothelin‐1 gene expression via protein kinase C pathway in human endothelial cells
Author(s) -
Wang Danny Ling,
Wung BeingSun,
Peng YiChing,
Wang Jaang Jung
Publication year - 1995
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.1041630220
Subject(s) - protein kinase c , calphostin c , microbiology and biotechnology , endothelin 1 , umbilical vein , gene expression , protein kinase a , messenger rna , secretion , calphostin , biology , chemistry , kinase , endocrinology , gene , in vitro , biochemistry , receptor
Vascular endothelial cells (ECs) are constantly subjected to mechanical strain due to relaxation and contraction of vessel walls. The effects of cyclical strain on endothelin‐1 (Et‐1) secretion and Et‐1 mRNA levels in human umbilical vein ECs were examined. Cultured ECs grown on a flexible membrane base were deformed by negative pressure (16 kPa at 60 cycles/min). Cells subjected to strain showed increased Et‐1 secretion (0.54 ng/hr/10 6 cells) compared with unstrained control cells (0.22 ng/hr/10 6 cells). Northern blot analysis of cells strained for 2 hours or longer demonstrated a sustained elevated Et‐1 mRNA level at more than double the level in unstrained controls. This strain‐induced ET‐1 mRNA level returned to its basal level 2 hours after the release of strain. Cells treated with actinomycin D before or during strain treatment showed no strain‐induced gene expression. Pretreatment of ECs with a protein kinase C (PKC) inhibitor, Calphostin C, strongly inhibited the strain‐induced Et‐1 gene expression. Pretreatment of ECs with cAMP or cGMP‐dependent protein kinase inhibitors (KT5720 or KT5823) only partially inhibited the increased Et‐1 mRNA levels in strain‐treated cells. EGTA strongly inhibited the Et‐1 gene expression. The intracellular calcium chelator BAPTA/AM also showed an inhibitory effect on Et‐1 mRNA levels. We conclude that mechanical strain can stimulate the secretion of Et‐1 from ECs by increasing Et‐1 mRNA levels via transcription, and that this gene induction is mediated predominantly via the PKC pathway and requires extracellular Ca 2+ . This strain‐induced Et‐1 gene expression in ECs may contribute to the regulation of vascular tone and structure in normal and pathological states of the cardiovascular system. © 1995 Wiley‐Liss, Inc.