Synthesis of tissue inhibitor of metalloproteinase-1 (TIMP-1) in rabbit aortic neointima after selective de-endothelialization

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      Altered TIMP-1 synthesis in the arterial wall may be important for the balance between metalloproteinases and their inhibitors, and thus contribute to dysregulated extracellular matrix metabolism in atherosclerotic lesions. To examine this, we cloned the rabbit TIMP-1 gene from aortic neointima, developed in response to a balloon-catheter induced de-endothelialization. The apparent homology of cDNA with TIMP-1 genes from several sources suggested that it is a rabbit form of TIMP-1. We examined the recombinant rabbit TIMP-1 expression in Escherichia coli using the pTrxFus expression system and the synthesis of the resulting soluble protein was confirmed by immunostaining with anti-TIMP-1. The TIMP-1 concentration in normal and de-endothelialized rabbit aortas was compared using Northern blot, Western blot and mRNA in situ hybridization techniques. We observed a significant increase of TIMP-1 expression in neointimal SMCs at both nucleic acid and protein levels, suggesting a role of TIMP-1 in injury-induced atherogenesis.


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        • Moore S
        • Alavi MZ
        Vessel wall matrix: an overview.
        in: Woodford F.P. Davignon J. Sniderman A. Proceedings of the 10th International Congress of Atherosclerosis. Elsevier, Amsterdam1995: 369
        • Li Z
        • Alavi MZ
        • Wasty F
        • Galis Z
        • Moore S
        Proteoglycan synthesis by the neointimal smooth muscle cells cultured from rabbit aortic explants following de-endothelialization.
        Pathobiology. 1993; 61: 89
        • Galis Z
        • Alavi MZ
        • Moore S
        Co-localization of apolipoprotein B and chondroitin sulfate proteoglycans in an injury model of atherosclerosis.
        Am J Pathol. 1993; 142: 1432
        • Camejo G
        • Fager G
        • Rosengren B
        • Hurt CE
        • Bondjers G
        Binding of low density lipoproteins by proteoglycans synthesized by proliferating and quiescent human arterial smooth muscle cells.
        J Biol Chem. 1993; 268: 14131
        • Libby P
        Molecular bases of the acute coronary syndromes.
        Circulation. 1995; 91: 2844
        • Woessner JF
        The family of matrix metalloproteinases.
        Ann NY Acad Sci. 1994; 732: 11
        • Reynolds JJ
        • Hembry RM
        • Meikle MC
        Connective tissue degradation in health and peridontal disease and the roles of matrix metalloproteinases and their natural inhibitors.
        Adv Dent Res. 1994; 8: 312
        • Krane SK
        Clinical importance of metalloproteinases and their inhibitors.
        Ann NY Acad Sci. 1994; 732: 1
        • Docherty AJP
        • Lyons A
        • Smith BJ
        • Wright EM
        • Stephens PE
        • Harris TJR
        • Murphy G
        • Reynolds JJ
        Sequence of human TIMP and its identity to erythroid-potentiating activity.
        Nature. 1985; 318: 66
        • Denhardt DT
        • Feng B
        • Edwards DR
        • Cocuzzi ET
        • Malyankar UM
        Tissue inhibitor of metalloproteinases (TIMP, aka EPA): structure, control of expression and biological functions.
        Pharma Ther. 1993; 59: 329
        • Edwards DR
        • Rocheleau H
        • Sharma RR
        • Wills AJ
        • Cowie A
        • Hassell JA
        • Heath JK
        Involvement of AP1 and PEA3 binding sites in the regulation of murine tissue inhibitor of metalloproteinases-1 transcription.
        Biochim Biophys Acta. 1992; 117: 41
        • Roswit WT
        • McCourt DW
        • Partridge NC
        • Jeffery JJ
        Purification and sequence analysis of two rat tissue inhibitors of metalloproteinaess.
        Arch Biochem Biophys. 1992; 292: 402
        • Smith GW
        • Goetz TL
        • Anthony RL
        • Smith MF
        Molecular cloning of an ovine ovarian tissue inhibitor of metalloproteinases: ontogeny of messenger ribonucleic acid expression and in situ localization within pre-ovulatory follicles and luteal tissue.
        Endocrinology. 1994; 134: 344
        • Yang TT
        • Hawkes SP
        Role of the 21-kDa protein TIMP-3 in oncogenic transformation of cultured chicken embryo fibroblasts.
        in: Proc Natl Acad Sci USA. 89. 1992: 10676
        • Tilson MD
        • Reilly JM
        • Brophy CM
        • Webster EL
        • Barnett TR
        Expression and sequence of the gene for tissue inhibitor of metalloproteinases in patients with abdominal aortic aneurysms.
        J Vasc Surg. 1993; 18: 266
        • Docherty AJP
        • Murphy G
        The tissue metalloproteinase family and the inhibitor TIMP: a study using cDNA and recombinant proteins.
        Ann Rheum Dis. 1990; 49: 469
        • Huebner K
        • Isobe M
        • Gasson JC
        • Golde DW
        • Croce CM
        Localization of the gene encoding human erythroid-potentiating activity to chromosome region Xp11.1-Xp11.4.
        Am J Hum Genet. 1986; 38: 819
        • Okada A
        • Garnier J-M
        • Vicarie S
        • Basset P
        Cloning of the cDNA encoding rat tissue inhibitor of metalloproteinase-1, amino acid com parisien with other TIMPs, and gene expression in rat tissues.
        Gene. 1994; 147: 301
        • DeClerck YA
        Purification and characterization of collagenase inhibitor produced by bovine vascular smooth muscle cells.
        Arch Biochem Biophys. 1988; 265: 28
        • Galis ZS
        • Sukhova GK
        • Lark MW
        • Libby P
        Increased expression of matrix metalloproteinases and matrix degrading activity in vulnerable regions of human atherosclerotic plaques.
        J Clin Invest. 1994; 94: 2493
        • Alavi MZ
        • Moore S
        Kinetics of low density lipoprotein interaction with rabbit aortic wall following balloon catheter de-endothelialization.
        Arteriosclerosis. 1984; 4: 395
        • Chomcynski P
        • Sacchi N
        Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction.
        Anal Biochem. 1987; 162: 156
        • Bilder GE
        • Kasiewski CJ
        • Costello RJ
        • Hodge TG
        • Perrone MH
        Electrode cuff-induced changes in DNA and PDGF gene expression in the rat carotid artery.
        Atherosclerosis. 1993; 100: 103
        • Chomcynski P
        A reagent for the single-step simultaneous isolation of RNA, DNA and protein from cell and tissue samples.
        Biotechniques. 1993; 15: 532
        • Galis ZS
        • Ghitescu LD
        • Li Z
        • Alavi MZ
        • Moore S
        Distributation of endogenous apoprotein B-containing lipoproteins in normal and injured aortas of normoc- holesterolemic rabbits.
        Lab Invest. 1992; 66: 624
        • Murphy G
        • Koklitis P
        • Carne AF
        Dissociation of tissue inhibitor of metalloproteinases from enzyme complexes yields fully active inhibitor.
        Biochem J. 1989; 261: 1031
        • O'Shea M
        • Willenbrock F
        • Williamson RA
        • Cockett MI
        • Freedman RB
        • Reynolds JJ
        • Docherty AJP
        • Murphy G
        Site-directed mutation that alter the inhibitory activity of the tissue inhibitor of metalloproteinases-1: importance of the N-terminal region between cysteine 3 and cysteine 13.
        Biochemistry. 1992; 31: 10146
        • Freudenstein J
        • Wagner S
        • Luck RM
        • Eispaniere R
        • Scheit KH
        mRNA of bovine tissue inhibitor of metalloproteinases: sequence and expression in bovine ovarian tissue.
        Biochem Biophys Res Commun. 1990; 171: 250
        • Willenbrock F
        • Murphy G
        Structure-function relationships in the tissue inhibitors of metalloproteinases.
        Am J Respir Crit Care Med. 1994; 150: S165
        • Horowitz S
        • Dafni N
        • Shapiro DL
        • Holm BA
        • Notter RH
        • Quille DJ
        Hyperoxic exposure alters gene expression in the lung: induction of the tissue inhibitor of metalloproteinase mRNA and other mRNAs.
        J Biol Chem. 1989; 264: 7092
        • Campbell CE
        • Flenniken AM
        • Skup D
        • William BRG
        Identification of a serum and phorbol-ester responsive element in the murine tissue inhibitor of metalloproteinase gene.
        J Biol Chem. 1991; 266: 7199
        • Leco KJ
        • Hayden LJ
        • Sharma RR
        • Rocheleau H
        • Greenberg AH
        • Edwards DR
        Differentiatial regulation of TIMP-1 and TIMP-2 mRNA expression in normal and Ha-ras-transformed murine fibroblasts.
        Gene. 1992; 117: 209
        • Lotz M
        • Guerne PA
        Interleukin-6 induces the synthesis of tissue inhibitor of metalloproteinase-1/erythroid potentiating activity (TIMP-1/EPA).
        J Biol Chem. 1991; 266: 2017
        • Shapiro SD
        • Doyle GAR
        • Ley TJ
        • Parks WC
        • Welgus HG
        Molecular mechanisms regulating the production of collagenase and tissue inhibitor of metalloproteinase in U937 cells: evidence for involvement of delayed transcritpional activation and enhanced mRNA stability.
        Biochemistry. 1993; 32: 4286
        • Schwartz SM
        • deBlois D
        • O'Brien ERM
        The intima: soil for atherosclerosis and restenosis.
        Circ Res. 1995; 77: 445
        • Kordula T
        • Guttgemann I
        • Rose-John S
        • Boeb E
        • Osthues A
        • Tschesche H
        • Koj A
        • Heinrich PC
        • Graeve L
        Synthesis of tissue inhibitor of metalloproteinase-1 (TIMP-1) in human hepatoma cells (HepG2).
        FEBS Lett. 1992; 313: 143
        • Galis ZS
        • Muszynski M
        • Sukhova GK
        • Simon-Morrissey E
        • Libby P
        Enhanced expression of vascular matrix metalloproteinases induced in vitro by cytokines and in regions of human atherosclerotic lesions.
        Ann NY Acad Sci. 1994; 748: 501
        • Leco KJ
        • Khokha R
        • Pavloff N
        • Hawkes SP
        • Edwards DR
        Tissue inhibtor of metalloproteinases-3 is an extracellular matrix-associated protein with a distinctive pattern of expression in mouse cells and tissues.
        J Biol Chem. 1994; 269: 9352
        • Dean DD
        • Martel-Pelletier J
        • Pelletier J-P
        • Howell DS
        • Woessner Jr., JF
        Evidence for metalloproteinase and metalloproteinase inhibitor imbalance in human osteoarthritic cartilage.
        J Clin Invest. 1989; 84: 678
        • Khokha R
        • Waterhouse P
        • Yagel S
        • Lala PK
        • Overall CM
        • Norton G
        • Denhardt DT
        Antisense RNA-induced reduction in murine TIMP levels confers oncogenicity on Swiss 3T3 cells.
        Science. 1989; 43: 947
        • Lindsay CK
        • Thorgeirsson UP
        Localization of messenger RNA for TIMP-1 and type IV collagenases/gelatinases in monkey hepatocellular carcinomas.
        Clin Experimen Metast. 1995; 13: 381
        • Stetler-Stevenson WG
        • Liotta LA
        • Kleiner Jr, DE
        Extracellular matrix 6: role of matrix metalloproteinases in tumor invasion and metastasis.
        FASEB J. 1993; 7: 1434
        • Zempo O
        • Kenagy RD
        • Au YPT
        • Bendeck M
        • Clowes MM
        • Reidy MA
        • Clowes AW
        Matrix metalloproteinases of vascular wall cells are increased in balloon-injured rat carotid artery.
        J Vasc Surg. 1994; 20: 209
        • Hayakawa T
        • Yamashita K
        • Tanzawa K
        • Uchijima E
        • Iwata K
        Growth promoting activity of tissue inhibitor of metalloproteinase-1 for a wide range of cells: a possible new growth factor in serum.
        FEBS Lett. 1992; 298: 29
        • Stricklin GP
        • Nanney LB
        Immunolocalization of collagenase and TIMP in healing human burn wounds.
        J Invest Dermatol. 1994; 103: 488