Human-derived vascular smooth muscle cells produce angiotensin II by changing to the synthetic phenotype.
We investigated whether vascular smooth muscle cells (VSMC)-derived from human produce angiotensin (Ang) II upon change from the contractile phenotype to the synthetic phenotype by incubation with fibronectin (FN). Expression of alpha-smooth muscle (SM) actin, apparent in the contractile phenotype, was decreased by FN. Expressions of matrix Gla and osteopontin, apparent in the synthetic phenotype, were increased by FN. Ang II measured by radioimmunoassay (RIA) was significantly increased in human VSMC by FN. Expression of mRNAs for Ang II-generating proteases cathepsin D, cathepsin G, ACE, and chymase was increased by FN. Expressions of cathepsin D and cathepsin G proteins were also increased by FN. Ang I-generating activity, which was inhibited by an aspartyl protease inhibitor pepstatin A, was readily detected in the conditioned medium from human VSMC. Antisense oligodeoxynucleotides (ODNs) that hybridize with cathepsin D and cathepsin G significantly inhibited FN-increased Ang II in conditioned medium and cell extracts. In VSMC conditioned medium, FN-induced elevation of Ang II was significantly inhibited by temocapril but not by chymostatin. Ang II type 1 receptor antagonist CV11974 completely, and antisense cathepsin D and cathepsin G ODNs partially inhibited the FN-stimulated growth of human VSMC. These results indicate that the change of homogeneous cultures of human VSMC from the contractile to the synthetic phenotype sequentially increases expression of proteases cathepsin D, cathepsin G, and ACE, production of Ang II and productions of growth factors, culminating in VSMC proliferation. These findings implicate a new mechanism for the pathogenesis of human vascular proliferative diseases.
Hu
Wen-Yang
WY
Second Department of Internal Medicine, Nihon University School of Medicine, Tokyo, Japan.
Fukuda
Noboru
N
Ikeda
Yukihiro
Y
Suzuki
Ryo
R
Tahira
Yosiko
Y
Takagi
Hiroto
H
Matsumoto
Koichi
K
Kanmatsuse
Katsuo
K
Mugishima
Hideo
H
D000803:Angiotensin I / Q000096:biosynthesis; D000804:Angiotensin II / Q000096:biosynthesis*; D000806:Angiotensin-Converting Enzyme Inhibitors / Q000494:pharmacology; D002402:Cathepsin D / Q000235:genetics; D056649:Cathepsin G; D002403:Cathepsins / Q000235:genetics; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D005353:Fibronectins / Q000494:pharmacology; D006801:Humans; D009131:Muscle, Smooth, Vascular / Q000187:drug effects / Q000378:metabolism* / Q000502:physiology; D032389:Myocytes, Smooth Muscle / Q000187:drug effects / Q000378:metabolism* / Q000502:physiology; D016376:Oligonucleotides, Antisense / Q000494:pharmacology; D009842:Oligopeptides / Q000494:pharmacology; D010641:Phenotype; D012697:Serine Endopeptidases; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology; D013841:Thiazepines / Q000494:pharmacology
D000806:Angiotensin-Converting Enzyme Inhibitors; D005353:Fibronectins; D016376:Oligonucleotides, Antisense; D009842:Oligopeptides; D015842:Serine Proteinase Inhibitors; D013841:Thiazepines; D000804:Angiotensin II; C055603:temocapril hydrochloride; D000803:Angiotensin I; C002101:chymostatin; D002403:Cathepsins; D012697:Serine Endopeptidases; C534094:CTSG protein, human; D056649:Cathepsin G; D002402:Cathepsin D