Abstract
Objective:
Protease-nexin 1 (PN-1) belongs to the serpin superfamily and behaves as a specific thrombin inhibitor in the pericellular environment. Little is known about PN-1 expression and its regulation in the vascular system. In this study, we examined the expression of functionally active PN-1 in vitro in rat aortic smooth muscle cells and in vivo in rat arterial media and its regulation in hypertensive rats.
Methods and results:
The vascular PN-1 formed specific covalent complexes with thrombin involving the catalytic site of the protease, and heparin increased the formation of these complexes. We also demonstrated PN-1 in rat arterial media by immunohistochemical staining. Moreover, we examined in vivo vascular expression of PN-1 in a model of chronic hypertension induced by long-term administration of N(G)-nitro-L-arginine methyl ester (L-NAME). Marked increases in PN-1 mRNA (3-fold) and protein (2-fold) were observed after 2 months of hypertension. Increased expression of PN-1 in the vascular wall was associated with an increase in the formation of complexes between radiolabeled-thrombin and PN-1, indicating that PN-1 was functional.
Conclusions:
PN-1 may thus participate in the mechanisms that regulate thrombin activity in the vessel wall.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Administration, Oral
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Amyloid beta-Protein Precursor
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Angiotensin II / pharmacology
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Animals
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Aorta / metabolism*
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Aorta / pathology
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Calcium / metabolism
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Carrier Proteins / biosynthesis*
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Carrier Proteins / immunology
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Carrier Proteins / pharmacology
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Carrier Proteins / physiology*
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Cell Division / drug effects
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Cells, Cultured
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Humans
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Hypertension / chemically induced*
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Hypertension / metabolism*
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Immunohistochemistry
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Male
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Muscle, Smooth, Vascular / chemistry*
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Muscle, Smooth, Vascular / drug effects
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Muscle, Smooth, Vascular / metabolism
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Muscle, Smooth, Vascular / pathology
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NG-Nitroarginine Methyl Ester / administration & dosage*
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Nitric Oxide Donors / pharmacology
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Protease Nexins
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Rats
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Rats, Wistar
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Receptors, Cell Surface
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Recombinant Proteins / pharmacology
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Serpin E2
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Serpins / biosynthesis*
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Serpins / immunology
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Serpins / pharmacology
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Serpins / physiology*
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Thrombin / antagonists & inhibitors
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Thrombin / pharmacology
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Up-Regulation / drug effects
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Up-Regulation / physiology*
Substances
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Amyloid beta-Protein Precursor
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Carrier Proteins
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Nitric Oxide Donors
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Protease Nexins
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Receptors, Cell Surface
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Recombinant Proteins
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SERPINE2 protein, human
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Serpin E2
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Serpins
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Angiotensin II
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Thrombin
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Calcium
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NG-Nitroarginine Methyl Ester