KATP channel blocking actions of quaternary ions play no role in their antiproliferative action on mouse leukaemia and rat vascular smooth muscle cells in vitro.

1. The aim of the present study was to investigate the possibility that, in the two cell lines examined, alterations in cell growth caused by lipophilic quaternary ions may involve KATP channels. We examined the effect of tetraphenylphosphonium (TPP), tetraphenylboron (TPB), rhodamine 123, dequalinium chloride (DECA) and the non-quaternary ion cisplatin on the proliferation of L1210 mouse leukaemia cells and rat smooth muscle cells in vitro. The KATP channel opener levcromakalim (LKM) and the KATP channel antagonist glibenclamide were also tested. 2. From growth-inhibition studies, the rank order of potency (based on pIC50 values) using L1210 leukaemia cells was: DECA (6.61) > cisplatin (6.09) = rhodamine 123 (6.01) > TPP (5.61) > TPB (4.25). Levcromakalim and glibenclamide were found to be inactive at the maximum concentrations used (100 mumol/L). A different rank order of potency was obtained in rat aortic smooth muscle cells: cisplatin (6.33) > DECA (5.67) > TPP (4.96) > rhodamine 123 (4.1). Tetraphenylboron (30 mumol/L), LKM (100 mumol/L) and glibenclamide (100 mumol/L) were found to be inactive. 3. When the negatively charged TPB (30 mumol/L) was combined with some of the active agents, the potency of the active agents was increased. Thus, in L1210 cells, rhodamine 123, DECA and TPP were all more potent at inhibiting cell growth in the presence of TPB. Tetraphenylboron had no effect on cisplatin in this cell line. In rat smooth muscle cells, TPB (30 mumol/L) potentiated the effect of rhodamine 123 but had no effect on the actions of cisplatin, DECA or TPP. 4. In functional studies, rhodamine 123 was a weak antagonist of the vasorelaxant responses to the KATP channel opener LKM in the porcine right circumflex artery in vitro. The pKB value obtained for rhodamine 123 at 100 mumol/L was 4.95. Dequalinium chloride was inactive. 5. We found no correlation between the actions of the compounds tested to antagonise KATP channels and their ability to inhibit cell proliferation. In addition, compounds known to regulate KATP channel activity failed to influence proliferative rates. These results suggest that KATP channels are not involved in the antiproliferative action of TPP and other quaternary ions in the two cell lines studied.

Regulation by protein kinase C of transforming growth factor-beta 1 action on the proliferation of vascular smooth muscle cells from spontaneously hypertensive rats.

1. This study examined the role of protein kinase C (PKC) on the action of transforming growth factor-beta 1 (TGF-beta 1) to regulate the proliferation of vascular smooth muscle cells (VSMC) isolated from the aorta of the spontaneously hypertensive rat (SHR). 2. Down-regulation of PKC by prolonged exposure to phorbol 12-myristate 13-acetate (PMA) completely inhibited the ability of TGF-beta 1 to potentiate epidermal growth factor-stimulated proliferation of VSMC. 3. In contrast, the inhibitory effect of TGF-beta 1 on serum-stimulated proliferation of VSMC was not altered by PMA action. 4. These results suggest that PKC-dependent signalling pathways involved in the regulation of growth by TGF-beta 1 may be important in any proliferative component of vascular hypertrophy that develops in the SHR.

Regulation and interactions of transforming growth factor-beta with cardiovascular cells: implications for development and disease.

1. Transforming growth factors-beta (TGF-beta) are multifunctional proteins that regulate cell growth, differentiation, migration and extracellular matrix production and have an important role in embryonic development and tissue remodelling. 2. The diverse biological actions of TGF-beta are elicited following their interaction with type I and type II TGF-beta receptors, both of which are transmembrane serine/threonine kinases, suggesting an important role for protein phosphorylation in the mechanism of action of these cytokines on the growth of cells and their extracellular environment. 3. Alterations in TGF-beta gene expression and action in various cell types associated with the cardiovascular system may contribute to the pathophysiology of a number of diseases, such as hypertension, atherosclerosis and restenosis, as well as the development of cardiac abnormalities.

TGF-beta: receptors and cell cycle arrest.

Transforming growth factor-beta (TGF-beta) is the prototypic member of a superfamily of proteins important in normal growth and development. The recent molecular cloning and characterization of TGF-beta receptors represents a major advance in our understanding of the mechanism of action of this cytokine. These studies have revealed that TGF-beta elicits its diverse biological responses following the formation of a heteromeric complex involving two distantly related transmembrane serine/threonine kinases, both of which are required for signalling. Furthermore, there has been very rapid progress in the identification of key components that regulate the cell cycle, including the cyclin-dependent kinases (CDKs) and their partners, the cyclins. Perhaps the most significant development has been the isolation of a family of proteins that bind to and inactivate CDKs. The ability of TGF-beta to regulate the action of these CDK inhibitors results in growth arrest of mammalian cells in G1.

Developmental regulation of transforming growth factor-beta 1 responses and vascular smooth muscle growth in spontaneously hypertensive rats.

OBJECTIVE: To investigate the developmental regulation of transforming growth factor-beta 1 (TGF-beta 1) action and the expression of TGF-beta receptors in vascular smooth muscle cells (VSMC) isolated from spontaneously hypertensive rats (SHR). DESIGN: TGF-beta 1 effects on proliferation and expression of TGF-beta receptor subtypes were compared in VSMC prepared from SHR, Wistar-Kyoto (WKY) and Sprague-Dawley rats of different ages. METHODS: TGF-beta 1 effects on platelet-derived growth factor-BB (PDGF-BB)-stimulated proliferation were examined in VSMC isolated from SHR, WKY and Sprague-Dawley rats aged 1, 4 and 12 weeks, and from renal hypertensive WKY rats. TGF-beta receptors on the surface of VSMC were identified by affinity labelling, followed by immunoprecipitation with TGF-beta receptor antibodies; complexes were then analysed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis. RESULTS: TGF-beta 1 inhibited by 60-80% PDGF-BB-stimulated proliferation of VSMC from 1-week-old SHR. In 4-week-old SHR, VSMC were resistant to the antiproliferative action of TGF-beta 1, whereas the mitogenic activity of PDGF-BB was increased approximately 150% by TGF-beta 1 in VSMC from 12-week-old SHR. In contrast, TGF-beta 1 inhibited by 10-50% PDGF-BB-stimulated proliferation of VSMC from age-matched WKY and Sprague-Dawley rats. TGF-beta isoforms (TGF-beta 1, -beta 2, -beta 3) all elicited similar growth responses in VSMC from SHR and WKY rats of different ages. Hypertension per se did not alter TGF-beta 1 effects on proliferation, as TGF-beta 1 inhibited by 30-40% growth factor action on VSMC from control, uni-nephrectomized and one-kidney one-clip hypertensive WKY rats. The type I, II and III TGF-beta receptors were expressed on the surface of VSMC isolated from SHR of different ages. CONCLUSION: Alterations in TGF-beta 1 responses, which become evident in VSMC from 4-week-old SHR and are most prominent at 12 weeks, may be important in the development of vascular hypertrophy in this rat strain.

Expression of growth factor receptors in arterial smooth muscle cells. Dependency on cell phenotype and serum factors.

The effects of modulation of rabbit aortic smooth muscle cells (SMCs) from the 'contractile' phenotype on surface membrane receptors binding epidermal growth factor (EGF), basic fibroblast growth factor (bFGF) and platelet-derived growth factor-BB (PDGF-BB), as well as their responsiveness to these growth factors was investigated in cell culture. Cells predominantly of the 'contractile' phenotype expressed low numbers of high affinity EGF and bFGF receptors (EGFr: 1.09 +/- 0.18 fmol/10(6) cells; bFGFr: 0.32 +/- 0.07 fmol/10(6) cells). Upon modulation from the 'contractile' phenotype, the expression of these cell surface receptors increased greatly: 8- and 11-fold with respect to EGF and bFGF receptors. Cell surface receptors binding [125I]-PDGF-BB were largely unaltered. The elevated bFGF receptor number appeared dependent on SMC modulation from the 'contractile' phenotype and serum; the latter factor did not influence EGF receptor numbers. In both instances the increase in receptor numbers was independent of the proliferation status of the cells. Cells expressing high levels of the growth factor receptors also rapidly entered the cell cycle, proliferated, and exhibited growth factor-specific changes in shape in the presence of these growth factors. Because the effects on growth factor receptor numbers were observed in confluent cells, such alterations, are likely to play a significant role in vessel remodelling following balloon catheter angioplasty, in atherosclerotic vessels and the vascular hypertrophy associated with hypertension.

Effects of nerve growth factor on sympathetic neuron development in normal and limbless chick embryos.

Wing bud removal in chick embryos has been shown to affect the generation of sympathetic neurons prior to the normal period of limb innervation [Saltis and Rush (1995) J. auton. nerv. Sys., 51, 117-127.]. Pyknotic activity occurred earlier within the peripherally deprived ganglion, suggesting that a precocious cell death of dividing sympathoblasts led to the reduced neuronal population. We have now sought to test whether the effect of limb bud extirpation can be overcome by the administration of nerve growth factor (NGF). Specifically, the peripherally deprived ganglion has been examined for mitotic activity and total neuronal numbers. In brachial ganglia from the operated side, neuron numbers decreased by 67% by embryonic day (E) 13, but by only 28% when NGF was administered from E9. Ganglia on the unoperated side were unaffected by the NGF treatment. In contrast, in embryos receiving NGF from E5 to E9, neuron numbers in the ganglia increased by more than 100%, on both the intact and operated side. This increase was accompanied by a greater proportion of 3H-thymidine-labelled neurons. We therefore conclude that NGF, in addition to its previously described role of preventing naturally occurring neuron death, can also affect the generation of sympathetic neurons. This ability of NGF to affect gangliogenesis is most likely achieved by increasing the survival of dividing neuroblasts, although a direct effect on mitosis has not been excluded.

Glucocorticoids but not mineralocorticoids modulate endothelin-1 and angiotensin II binding in SHR vascular smooth muscle cells.

Both glucocorticoids and mineralocorticoids are involved in circulatory homoeostasis and blood pressure control. In recent years direct effects of both steroid classes on vascular smooth muscle cells (VSMC) have been reported. We have thus examined the effects of RU 28362, a pure glucocorticoid agonist, and aldosterone, the physiologic mineralocorticoid, on the binding to VSMC from spontaneously hypertensive rats (SHR) of two key vasoactive peptides, endothelin-1 and angiotensin II. Binding of angiotensin II rose, and that of endothelin-1 declined, in a time- and dose-dependent fashion with maximal effects observed at 24 h and half-maximal effects for each at 2-3 nM RU 28362. Scatchard analysis showed that for both endothelin-1 and angiotensin II, RU 28362 alters receptor number but not affinity; competition studies with receptor-selective ligands (BQ123, S6C, DuP753 and PD123319) show that glucocorticoids specifically elevate (X2) AT-1 receptors and specifically lower (to approximately 30%) levels of ETA receptors. Treatment of VSMC with the antiglucocorticoid RU 38486 reversed the effect of glucocorticoids on endothelin-1 and angiotensin II binding, confirming the Type II (glucocorticoid) receptor mediated effect of the glucocorticoids. Aldosterone (100 nM) also lowers endothelin-1 binding and increases angiotensin II binding in VSMC; that this effect reflects aldosterone occupancy of classical glucocorticoid receptors is shown by the blockade of the aldosterone effect by an equal concentration (100 nM) of RU 38486--i.e. there is no evidence for an action of aldosterone via mineralocorticoid receptors. We interpret our results as evidence for a complex modulation of receptors for vasoactive peptides in VSMC by glucocorticoid but not mineralocorticoid hormones.

The development of normal and peripherally deprived sympathetic neurons in the chick.

Sympathetic neuron numbers in four brachial paravertebral ganglia (segments 13-16) were quantified during normal development and following wing bud removal. In ganglia with an intact peripheral field, different developmental profiles were observed. In particular, a period of increasing neuron numbers was seen in some, but not all ganglia from embryonic day 10. Similarly, a period of declining neuron numbers was present only in two ganglia, and this occurred at a time when sympathetic nerve fibres were detectable in the wing. Peak pyknotic activity occurred well before nerve fibres entered the wing. Thus, the bulk of the cell death is unrelated to peripheral innervation. Wing bud removal at embryonic day 3 reduced neuron number by as much as 69% at embryonic day 20 compared to the number in the corresponding ganglia on the intact side. The most obvious effect was the failure to generate the full complement of neurons. This could be explained by a shift in pyknotic activity to a period of 1-2 days earlier than seen on the intact side. We therefore conclude that the developing wing influences the size of the sympathetic population both before and during the period of peripheral innervation.

Transforming growth factor-beta 1 and the development of vascular hypertrophy in hypertension.

Transforming growth factor-beta 1 (TGF-beta 1) is a multifunctional cytokine capable of regulating the growth and differentiation of many cell types, as well as regulating their environment in the blood vessel wall. Its production by endothelium and/or vascular smooth muscle is stimulated by biophysical forces, growth factors and also vasoconstrictors. In hypertension TGF-beta 1 gene transcription is most likely elevated by a combination of physical and chemical stimuli with the cytokine acting to increase the production of extracellular matrix proteins or to modulate smooth muscle cellular growth, producing hypertrophy, polyploidy or proliferation. With respect to the latter, TGF-beta 1 potentiates the proliferative effects of many receptor tyrosine kinase-activating growth factors in vascular smooth muscle from SHR, but inhibits such proliferation in WKY smooth muscle. It also differentially affects collagen production by the two cell types. It is suggested that the augmented proliferative response in renal hypertensive SHR, compared to renal hypertensive WKY, is the consequence of these differential effects of TGF-beta 1 on smooth muscle cell proliferation. TGF-beta 1 is also likely to be a significant contributor to the development of vascular hypertrophy in genetic hypertension.

Transforming growth factor-beta 1 gene activation and growth of smooth muscle from hypertensive rats.

Cultured vascular smooth muscle cells derived from the spontaneously hypertensive rat (SHR) are known to replicate more rapidly than cells from the normotensive Wistar-Kyoto (WKY) rat. In this study we compared the responses of vascular smooth muscle cells from the two strains to transforming growth factor-beta 1 (TGF-beta 1) and evaluated its potential to account for the different growth properties of these cells in response to a number of vascular-derived growth factors. TGF-beta 1 potentiated the proliferative effects of epidermal growth factor, basic fibroblast growth factor, or the different isoforms of platelet-derived growth factor on vascular smooth muscle cells from SHR but inhibited growth factor-stimulated proliferation of vascular smooth muscle cells from WKY rats. These differential effects of TGF-beta 1 on proliferation could not be attributed to alterations in the expression of the type I, II, or III TGF-beta receptors but appeared more related to the ability of cells to autoinduce the TGF-beta 1 gene. TGF-beta 1 caused a time-dependent increase in its own mRNA levels in vascular smooth muscle cells of WKY rats but attenuated levels in vascular smooth muscle cells of SHR. This effect was specific to TGF-beta 1 autoinduction since similar elevations in TGF-beta 1 mRNA levels were observed when vascular smooth muscle cells from the two rat strains were exposed to phorbol myristate acetate, basic fibroblast growth factor, or platelet-derived growth factor-BB.(ABSTRACT TRUNCATED AT 250 WORDS)

Developmentally regulated transforming growth factor-beta 1 action on vascular smooth muscle growth in the SHR.

1. This study examined the effects of transforming growth factor-beta 1 (TGF-beta 1) on platelet-derived growth factor-BB (PDGF-BB)-stimulated proliferation of vascular smooth muscle cells (VSMC) isolated from aortic tissue of 1, 4 and 12 week old spontaneously hypertensive rats (SHR). 2. In 1 week old SHR, TGF-beta 1 inhibited by about 60% VSMC proliferation stimulated by PDGF-BB; this inhibitory action of TGF-beta 1 was absent in VSMC isolated from 4 week old, prehypertensive SHR. In contrast, TGF-beta 1 potentiated by about 125% the mitogenic activity of PDGF-BB in VSMC cultures from adult SHR. 3. Age-dependent alterations in the action of TGF-beta 1 suggests an important role for TGF-beta 1 in the development of vascular hypertrophy from adolescence onwards in the SHR.

Effect of transforming growth factor-beta 1 on platelet-derived growth factor receptor binding and gene expression in vascular smooth muscle cells from SHR and WKY rats.

1. This study examined the effects of transforming growth factor-beta 1 (TGF-beta 1) on platelet-derived growth factor-BB (PDGF-BB)-stimulated DNA synthesis, [125I]-PDGF-BB receptor binding and PDGF-beta receptor mRNA expression in vascular smooth muscle cells (VSMC) isolated from Wistar-Kyoto (WKY) rats and spontaneously hypertensive rats (SHR). 2. TGF-beta 1 inhibited by 40% DNA synthesis stimulated by PDGF-BB in VSMC from WKY rats but potentiated by 20% growth factor-stimulated DNA synthesis in VSMC from the SHR. 3. Since the difference in effect of TGF-beta 1 could not be attributed to differential regulation of [125I]-PDGF-BB binding activity and PDGF-beta receptor mRNA expression, it is suggested that alterations in intracellular signalling pathways may account for the differential effects of TGF-beta 1 on PDGF-BB-stimulated DNA synthesis in VSMC from SHR and WKY rats.

Differences in growth characteristics of vascular smooth muscle from spontaneously hypertensive and Wistar-Kyoto rats are growth factor dependent.

OBJECTIVE: To investigate whether the differences in the growth properties between vascular smooth muscle cells (VSMC) from spontaneously hypertensive rats (SHR) and normotensive Wistar-Kyoto (WKY) rats depend on the type and nature of the growth factor. DESIGN: The growth characteristics of VSMC from SHR and WKY rats were compared in the presence of different growth factors. These were related to growth factor receptor expression on the VSMC. METHODS: Growth rates, cell densities at which VSMC become quiescent and [3H]-thymidine incorporation into DNA were examined in VSMC from SHR and WKY rats following exposure to epidermal growth factor (EGF), basic fibroblast growth factor (bFGF) or the three isoforms of platelet-derived growth factor (PDGF-AA, PDGF-AB and PDGF-BB). Receptor expression (receptor binding and Northern analysis) for these growth factors was also examined on VSMC from the two strains. RESULTS: VSMC from SHR exposed to the individual growth factors exhibited growth rates higher than those from WKY rats. For EGF, bFGF, PDGF-AB and PDGF-BB there was no difference in the sensitivity of VSMC to stimulation of [3H]-thymidine incorporation between the strains. In contrast, VSMC from SHR exhibited significantly increased sensitivity to the mitogenic effects of PDGF-AA. VSMC from SHR exposed to PDGF-BB or EGF proliferated and then became quiescent at densities approximately 50 and 200% higher, respectively, than those from WKY rats. In contrast, in the presence of bFGF, proliferating VSMC from WKY rats became quiescent at densities approximately 20% higher than those from SHR. Scatchard analysis of [125I]-labelled growth factor binding to VSMC from the two strains revealed similar receptor numbers and dissociation constants for all growth factors. Steady-state messenger RNA levels for the different receptors were also similar and could not account for the enhanced growth rates of VSMC from SHR. CONCLUSION: The rate and magnitude of the proliferative response elicited in VSMC cultures from SHR and WKY rats is critically dependent on the nature of the growth factor stimulus.

Age-dependent alterations in vascular smooth muscle cell responsiveness to platelet-derived growth factor in genetic hypertension.

1. This study examined and compared the growth characteristics of vascular smooth muscle cells (VSMC) isolated from 4 and 12 week old spontaneously hypertensive rats (SHR) and Wistar-Kyoto (WKY) rats following stimulation with platelet-derived growth factor-BB (PDGF-BB). 2. Vascular smooth muscle cells from 4 week old SHR proliferated at a slower rate than VSMC isolated from 12 week old SHR (1.08 +/- 0.06/day vs 1.89 +/- 0.13/day respectively, P < 0.05). In contrast, there was no difference in the proliferation of VSMC from 4 and 12 week WKY rats (0.62 +/- 0.06/day vs 0.82 +/- 0.13/day, respectively, P > 0.05). 3. The cell density at which VSMC from 4 and 12 week old SHR become refractory to the mitogenic effects of PDGF-BB was similar and approximately two-fold greater than VSMC from age-matched WKY rats. 4. This study concludes that there is an age-dependent, differential and specific upregulation of growth rate mechanisms in VSMC from SHR which enhance proliferation in response to PDGF-BB.

Vascular smooth muscle cell proliferation in SHR and WKY rats: evidence for specific differences in growth inhibitory regulatory mechanisms.

1. This study examined and compared the actions of transforming growth factor-beta 1 (TGF-beta 1), heparin, dexamethasone and interferon-gamma on platelet-derived growth factor-BB (PDGF-BB)-stimulated proliferation of vascular smooth muscle cells (VSMC) from normotensive, Wistar-Kyoto (WKY) rats and spontaneously hypertensive rats (SHR). 2. Heparin, dexamethasone and interferon-gamma all inhibited VSMC proliferation stimulated by PDGF-BB in both SHR and WKY rats. There was no difference (P > 0.05) in their inhibitory effects, which varied between 40 and 85% for the different agents. 3. Similarly, TGF-beta 1 inhibited PDGF-BB-stimulated VSMC proliferation in WKY rats by approximately 50%. In contrast, TGF-beta 1 potentiated growth factor action on cell proliferation in the SHR by approximately 40%. 4. Specific TGF-beta 1-stimulated regulatory mechanisms involved in the inhibition of proliferation are absent in SHR and this defect may contribute to the vascular hypertrophy which is apparent in genetic hypertension.

Dihydropyridine Ca2+ channel antagonists inhibit the salvage pathway for DNA synthesis in human vascular smooth muscle cells.

We examined the mechanisms by which Ca2+ channel antagonists inhibit the growth of smooth muscle cells by determining their effect on epidermal growth factor (EGF)-stimulated (i) induction of the early signalling gene, c-fos, (ii) incorporation of [3H]thymidine into cells as a measure of DNA synthesis, and (iii) increase in cell number. Verapamil, diltiazem, and the dihydropyridines felodipine, MDL 72892 A-15 (MDL) and nisoldipine had no effect on EGF-stimulated c-fos mRNA induction. Furthermore, only small inhibitory effects were observed on EGF-stimulated increases in cell number; felodipine, MDL, and nisoldipine at 0.3 microM inhibited EGF-stimulated cell proliferation by 9, 11, and 15%, respectively. In contrast, the dihydropyridine Ca2+ channel antagonists were found to be potent inhibitors of [3H]thymidine incorporation suggesting that they inhibit DNA synthesis. However, further examination revealed that the potent effects of dihydropyridine Ca2+ channel antagonists on [3H]thymidine incorporation were due not to an effect on incorporation of [3H]thymidine into DNA, but to a marked inhibitory effect on the cellular uptake of [3H]thymidine. Thus, we conclude that the small antiproliferative effects of the dihydropyridine antagonists are predominantly due to their ability to inhibit the activity of the salvage pathway for thymidylate synthesis in human vascular smooth muscle cells.

TGF-beta 1 potentiates growth factor-stimulated proliferation of vascular smooth muscle cells in genetic hypertension.

We have examined the interactions between transforming growth factor-beta 1 (TGF-beta 1) and epidermal growth factor (EGF), basic fibroblast growth factor (bFGF), or platelet-derived growth factor (PDGF) isoforms PDGF-AB and PDGF-BB on the proliferation of vascular smooth muscle cells isolated from the spontaneously hypertensive rat. TGF-beta 1 alone stimulated [3H]thymidine incorporation approximately twofold without a corresponding increase in cell number. In combination, TGF-beta 1 action was synergistic in further stimulating both DNA synthesis and cell proliferation 100-300% above the responses elicited by each growth factor. To gain further insight into the mechanism responsible for this potentiation, we examined the interaction between TGF-beta 1 and EGF. The synergistic interaction between TGF-beta 1 and EGF on DNA synthesis was independent of initial cell density. This effect of TGF-beta 1 was initiated early in the G1 phase of the cell cycle and did not appear to be mediated through the mobilization of Ca2+ or alterations in c-jun mRNA expression. However, in the presence of both TGF-beta 1 and EGF, there was a sustained elevation of c-myc mRNA levels over a 24-h period. These results suggest that TGF-beta 1 may interact with other growth factors in vivo to enhance their proliferative action on vascular smooth muscle of spontaneously hypertensive rats via mechanisms dependent on c-myc mRNA expression.

Vascular smooth muscle growth in genetic hypertension: evidence for multiple abnormalities in growth regulatory pathways.

OBJECTIVE: To gain insight into the mechanisms which contribute to the development of vascular hypertrophy in the spontaneously hypertensive rat (SHR). DESIGN: These experiments were performed under conditions which most closely mimic the growth of smooth muscle in blood vessels, i.e. once cell-cell contact has been achieved. METHODS: A comparison of the growth characteristics (growth rates and cell density at quiescence) of vascular smooth muscle cells (VSMC) from SHR and normotensive Wistar-Kyoto (WKY) rats. RESULTS: In the presence of foetal calf serum (1, 2.5, 5 and 10%), early passaged VSMC from SHR exhibited higher growth rates and reached higher densities at quiescence than VSMC from WKY rats. Accelerated growth rates could not be attributed to differences in cell-cell interactions. Also, growth rates and cell density at quiescence appear to be regulated by distinct mechanisms. Transforming growth factor-beta 1 (TGF-beta 1) caused an inhibition of serum-stimulated proliferation of confluent VSMC from WKY rats. In contrast, TGF-beta 1 had little, if any, inhibitory action upon the growth of VSMC from SHR. Scatchard analysis of 125I-TGF-beta 1 binding to VSMC from both strains yielded a single class of high affinity binding sites. CONCLUSIONS: VSMC from SHR exhibit enhanced proliferation, attain a higher cell density at quiescence and are less susceptible to growth inhibition by TGF-beta 1 than VSMC from WKY rats. All these characteristics of SHR VSMC may contribute to the development of vascular hypertrophy in this strain.

Differential regulation by transforming growth factor-beta 1 of platelet-derived growth factor-stimulated proliferation of vascular smooth muscle cells from SHR and WKY rats.

1. This study has examined and compared the time-course of action of transforming growth factor-beta 1 (TGF-beta 1) on platelet-derived growth factor-BB-stimulated proliferation of vascular smooth muscle cells isolated from normotensive Wistar-Kyoto (WKY) rats and spontaneously hypertensive rats (SHR). 2. Transforming growth factor-beta 1 inhibited vascular smooth muscle cell proliferation stimulated by platelet-derived growth factor-BB in WKY rats by approximately 60-75%. 3. In contrast, transforming growth factor-beta 1 potentiated an 8-35% growth factor action on cell proliferation in the SHR. 4. Defects in transforming growth factor-beta 1 action may be part of the molecular mechanism responsible for the development of vascular hypertrophy in the SHR.