Inhibition of rat arterial smooth muscle cell proliferation by heparin. II. In vitro studies.

Hoover, Richard L.; Rosenberg, Robert D.; Haering, W.A.; Karnovsky, Morris J.

doi:10.1161/01.res.47.4.578

Cited by 325 publications

(172 citation statements)

References 19 publications

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“…The negative charge of HA does not seem to be the sole factor responsible for its inhibitory effect, since chondroitin sulfate and dextran sulfate, which are more highly charged than HA, were ineffective. Heparin and heparan sulfate have been shown to inhibit proliferation of a variety of cell types (47)(48)(49)(50). In contrast to the inhibitory effect of HA described herein, the effect of heparin on smooth muscle cell proliferation is apparently mediated by specific cell surface receptors that recognize small oligosaccharides derived from heparin (51).…”

Section: Discussionmentioning

confidence: 72%

Hyaluronate inhibition of cell proliferation

Goldberg

Toole

1987

Arthritis & Rheumatism

174

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The proliferation of rabbit synovial cells, 3T3 cells, or simian virus-transformed 3T3 cells in cell culture was inhibited by the addition of hyaluronate to the culture medium. This effect was markedly dependent on the molecular weight and concentration of the hyaluronate. At the molecular weight and concentration of hyaluronate present in normal synovial fluid, proliferation was inhibited. At lower molecular weights or concentrations, as found in rheumatoid synovial fluid, hyaluronate was significantly less inhibitory. Thus, the changes in synovial fluid hyaluronate that are associated with arthropathies may contribute to a favorable environment for rheumatoid pannus expansion.The viscosity of synovial fluid from the rheumatoid joint is usually considerably less than that of normal synovial fluid, because of a decrease either in the degree of polymerization or in the concentration of hyaluronate (HA) in rheumatoid synovial fluid (1-4). A recent study suggests that the more important of these 2 factors is the decreased concentration of HA (4). Synovial cells, which are normally quiescent, are in constant contact with the HA of synovial fluid. In rheumatoid arthritis, these cells proliferate and con- In this study, we examined the effect of HA on proliferation of synovial and other cells in culture and found it to be inhibitory. However, our results indicate that the effect of HA on cell proliferation is markedly dependent on both the molecular weight and the concentration of HA in the cell culture medium. High molecular weight HA at high concentrations, as found in normal synovial fluid, is a potent inhibitor of cell division, whereas at lower molecular weights or at lower concentrations, as found in rheumatoid synovial fluid, it is less inhibitory and, in some cases, is even stimulatory . Therefore, alterations in either the concentration or the degree of polymerization of the HA of synovial fluid may contribute to the pathophysiology of rheumatoid arthritis by facilitating proliferation of the pannus. MATERIALS AND METHODSCells and culture procedures. Rabbit synovial fibroblast lines were obtained from Dr. C. Biswas (Tufts University School of Medicine, Boston, MA). These cells had been passaged in culture for approximately 2 years and had retained a constant morphology and the ability to produce large amounts of hyaluronate. The simian virus 40-transformed 3T3 (SV-3T3) and 3T3 cell lines were those used previously (5). All cells were maintained in Dulbecco's modified Eagle's medium (Gibco, Grand Island, NY) containing 10% fetal calf serum (Gibco), 100 units of penicillin per ml, 100 pg of streptomycin per ml, and 2.5 p g of fungizone per ml. The cells were grown and maintained in

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Section: Discussionmentioning

confidence: 72%

Hyaluronate inhibition of cell proliferation

Goldberg

Toole

1987

Arthritis & Rheumatism

174

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“…vations made in our laboratory over the past several years, which demonstrate that heparin inhibited the growth of SMC in vivo and in vitro (Clowes and Karuovsky, 1977;Guyton et al, 1980;Hoover et al, 1980;Castellot et at., 1981). In our previous structure-function studies (Castellot et al, 1984), we used heparin fragments and chemically modified commercial heparin to determine that the smallest antiproliferative fragment was a hexasaccharide, and that both N-and O-sulfates were important for maintaining the growth inhibitory capacity of heparin.…”

Section: Discussionmentioning

confidence: 99%

“…Earlier work from our laboratory demonstrated that heparin inhibited the proliferation of SMC in vivo and in vitro (Clowes and Karnovsky, 1977;Guyton et al, 1980;Hoover et al, 1980;Castellot et al, 1981). Both anticoagulant and non-anticoagulant heparin species were equally effective as SMC antiproliferative agents.…”

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confidence: 83%

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Structural determinants of the capacity of heparin to inhibit the proliferation of vascular smooth muscle cells. II. Evidence for a pentasaccharide sequence that contains a 3-O-sulfate group.

Castellot¹,

Choay²,

Lormeau³

et al. 1986

The Journal of Cell Biology

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155

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Abstract. Earlier work from our laboratory demonstrated that heparin inhibited the proliferation of vascular smooth muscle cells in vivo and in vitro. Both anticoagulant and non-anticoagulant heparin species were equally effective as antiproliferative agents. Previous structure-function studies indicated that hexasaccharide and larger fragments retained antiproliferative activity, whereas tetra-and disaccharides were inactive. These experiments also suggested that both Nand O-sulfates of heparin were necessary for growth inhibitory capacity. In this paper, we have further analyzed the structural determinants of the antiproliferative activity of heparin. These experiments were done using synthetically prepared and therefore chemically defined heparin oligosaccharides. We present evidence that a pentasaccharide fragment retains antiproliferative activity, and that the 3-O-sulfate on the internal glucosamine residue is critical for growth inhibitory capacity of the pentasaccharide. We also show that heparins obtained from different manufacturers differ significantly in their ability to suppress smooth muscle cell proliferation.T HE proliferation of vascular smooth muscle cells (SMC) ~ after endothelial injury is thought to be a key step in the pathogenesis of arteriosclerosis (Ross and Glomset, 1973). Although much attention has been focused on factors that stimulate SMC proliferation (Schwartz et al., 1982), very little is known about the mechanisms involved in maintaining these cells in a quiescent growth state in healthy blood vessels, and in re-establishing the quiescent growth state of SMC after their proliferative response to intimal injury.Earlier work from our laboratory demonstrated that heparin inhibited the proliferation of SMC in vivo and in vitro (Clowes and Karnovsky, 1977;Guyton et al., 1980;Hoover et al., 1980; Castellot et al., 1981). Both anticoagulant and non-anticoagulant heparin species were equally effective as SMC antiproliferative agents. These studies suggested both a pharmacological basis for the use ofhepafin after vessel injury, and a possible physiological role for heparin-like species as regulators of SMC growth in the vasculature.Previous structure-function studies (Castellot et al., 1984) indicated that hexasaccharide and larger fragments retained antiproliferative activity, whereas tetra-and disaccharides were inactive against SMC growth. These experiments also indicated that both the N-and O-sulfates of heparin were necessary for antiproliferative activity, although the N-sulfates could be replaced with acetyl groups without significant loss of activity. Both the hexasaccharide fragment and the Ndesulfated, re-N-acetylated heparin are devoid of anticoagulant activity (Kazatchkine et al., 1981;Oosta et al., 1981), thus providing a chemical basis for the difference between the antiproliferative and anticoagulant activities of this glycosaminoglycan.In this paper, we have further analyzed the structural determinants of the antiproliferative effect of heparin. These experiments were ...

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“…Azizkhan et al (1980) first reported that heparin induced expression of angiogenic substances in mast cells. Hoover et al (1980) subsequently demonstrated that heparin stimulated the growth of bovine aortic endothelial cells in vitro, but heparin was also demonstrated to inhibit the proliferation of fibroblasts and SMC. Potential biological roles of heparin in the process of angiogenesis has been mainly examined in relation to other growth factors, such as acidic fibroblast growth factor (aFGF) (Mueller et al 1989;Jesundason et al 2000), basic fibroblast growth factor (bFGF) (Yamashita et al 1992;Toriyama et al 1997) and vascular endothelial growth factor (VEGF) (Heaky et al 2000;Norrby 2000).…”

Section: Discussionmentioning

confidence: 99%

Heparin Accelerates Pulmonary Artery Development in Neonate Rabbits

Watanabe

Suzuki

Shizawa

et al. 2005

Tohoku J. Exp. Med.

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Pulmonary vascular resistance drops sharply within a few minutes after birth for the survival of neonates. A majority of this resistance is caused by "pulmonary vascular bed" or vessel lacking smooth muscle cells. Heparin is known to promote proliferation and development of endothelial cells and to subsequently decrease their overall vascular resistance, but its detailed features remained unknown. Therefore, in this study we treated neonatal rabbits with heparin, protamine (antagonist of heparin), or saline, and evaluated histopathological features of vascular endothelial cells using two different types of computer assisted image analysis, i.e., CAS200 and NIH image. These two systems detected the percentage of vascular endothelial area per fields (VA) and CD31-positive area per total area of tissue following subtraction of background stain. CD31 was used as an endothelial cell marker. Heparin treated rabbits were associated with significant decrement of pulmonary/systemic artery pressure (Pp/Ps) (21.0 ± 6.0%) compared to protamine (29.9 ± 6.1%) or saline (29.4 ± 3.0%) treated animals. The values of VA obtained by the two image analyses (CAS200 and NIH image) were significantly increased in heparin treated animals (38.4 ± 3.2% determined by CAS200 and 24.0 ± 1.3% by NIH image) compared to protamine (30.2 ± 3.9% and 19.2 ± 1.8%) or saline (33.2 ± 1.5% and 20.8 ± 3.8%) treated animals on 14th day of treatment. The present study indicates that heparin accelerates pulmonary vascular bed development probably by increasing the number and volume of endothelial cells, which subsequently contributes to the decrease in pulmonary vascular resistance. pulmonary artery vascular resistance; pulmonary vascular density; heparin; CD31; image analysis

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Inhibition of rat arterial smooth muscle cell proliferation by heparin. II. In vitro studies.

Cited by 325 publications

References 19 publications

Hyaluronate inhibition of cell proliferation

Hyaluronate inhibition of cell proliferation

Structural determinants of the capacity of heparin to inhibit the proliferation of vascular smooth muscle cells. II. Evidence for a pentasaccharide sequence that contains a 3-O-sulfate group.

Heparin Accelerates Pulmonary Artery Development in Neonate Rabbits

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