TY - JOUR. T1 - The mechanisms of uremic serum-induced expression of bone matrix proteins in bovine vascular smooth muscle cells. AU - Chen, N. X.. AU - Duan, D.. AU - ONeill, K. D.. AU - Wolisi, G. O.. AU - Koczman, J. J.. AU - LaClair, R.. AU - Moe, S. M.. PY - 2006/9/1. Y1 - 2006/9/1. N2 - We have previously found that uremic human serum upregulates RUNX2 in vascular smooth muscle cells (VSMCs), and that RUNX2 is upregulated in areas of vascular calcification in vivo. To confirm the role of RUNX2, we transiently transfected a dominant-negative RUNX2 (ΔRUNX2) construct in bovine vascular smooth muscle cells (BVSMCs). Blocking RUNX2 transcriptional activity significantly decreased uremic serum induced alkaline phosphatase (ALP) activity (268 ± 34 vs 188 ± 9.5 U/g protein, P , 0.05) and osteocalcin expression (172 ± 17 vs 125 ± 9 ODU, P , 0.05). To determine the mechanism by which uremic serum upregulates RUNX2, we examined cell signaling pathways. BVSMCs were incubated in the presence or ...
Hello. I need to culture bovine aortic smooth muscle cell. I do not know what medium should be used. What supplements or growth factors are required? In how much in amount? Please give me some help. Thanks in advance ...
Mouse Aortic Vascular Smooth Muscle Cells from Creative Bioarray are isolated from tissue of New Zealand White Rabbits. Rabbits Aortic Vascular Smooth Muscle Cells are grown in T25 tissue culture flasks pre-coated with gelatin-based solution for 0.5 hour and incubated in Creative Bioarrays Cell Culture Medium generally for 3-7 days. Cultures are then expanded. Prior to shipping, cells are detached from flasks and immediately cryo-preserved in vials. Each vial contains at least 1x10^6 cells per ml and is delivered frozen ...
The purpose of the present investigation was to explore the effects of well-defined flow conditions on the activity of tissue factor (TF) expressed on the surface of cultured rat vascular smooth muscle cells. Cells were cultured to confluence on Permanox brand slides and stimulated to express TF by a 90 min incubation with fresh growth medium containing 10 percent calf serum. The stimulated cells were then placed in a parallel plate flow chamber and perfused with Hanks Balanced Salt Solution containing factor VIIa, factor X (FX), and calcium. The chamber effluent was collected and assayed for factor Xa (FXa) and the steady-state flux of FXa was calculated. The flux values were 68.73, 94.81, 139.75, 138.19, 316.82, and 592.92 fmole/min/cm2 at wall shear rates of 10, 20, 40, 80, 320, and 1280 s−1 respectively. The FXa flux depended on the wall shear rate to a greater degree than predicted by classical mass transport theory. The flux at each shear rate was three to five times less than that ...
These findings point to a role of LTB4 in atherosclerosis and intimal hyperplasia, by identifying the vascular SMC as targets for this potent chemotactic molecule. The expression of the human BLT1 receptor on vascular SMC was demonstrated by immunohistochemical stainings of arterial samples, as well as in cultured human coronary SMC by Western blotting and RT-PCR. Together, these findings provide evidence that human vascular SMC express BLT1 receptors in vivo as well as in vitro, and they suggest that these cells may represent an additional target for LTB4.. Patch-clamp analysis and functional studies of SMC clarified that BLT1 receptors transduce a signal that leads to important functional responses in human vascular SMC. Membrane currents in human coronary artery SMC were increased significantly in the presence of either LTB4 or the selective BLT1 receptor partial agonist U75302. Also, another characteristic pharmacological feature of the BLT1 receptor (namely, its rapid desensitization by an ...
TY - JOUR. T1 - Nitric oxide reversibly inhibits the migration of cultured vascular smooth muscle cells. AU - Sarkar, Rajabrata. AU - Meinberg, Eric G.. AU - Stanley, James C.. AU - Gordon, R. David. AU - Webb, R Clinton. PY - 1996/1/1. Y1 - 1996/1/1. N2 - Augmentation of nitric oxide (NO) production in vivo decreases lesions in a variety of models of arterial injury, and inhibition of NO synthase exacerbates experimental intimal lesions. Both vascular smooth muscle cell (VSMC) proliferation and migration contribute to lesion formation. Although NO inhibit VSMC proliferation, its effects on VSMC migration are unknown. To test the hypothesis that NO inhibits VSMC migration independent of inhibition of proliferation, we examined migration of rat aortic VSMCs after wounding of a confluent culture in the presence of chemical donors of NO. Hydroxyurea was used to eliminate any confounding effect of NO on proliferation. Three NO donors, diethylamine NONOate, spermine NONOate, and S-nitrosoglutathione, ...
The concept of arterial SMC heterogeneity has gained wide acceptance in the last years.1 2 33 The distinct phenotypes of arterial SMCs have been mainly identified in vitro,4 5 6 7 8 9 10 11 12 13 14 15 16 suggesting that specific features of SMC populations arise and are maintained in the particular environment of cell culture. Hence, it was of interest to investigate whether in vitro SMC phenotypes are preserved when SMCs are placed back in an in vivo environment. For this purpose, we have implanted 2 SMC populations exhibiting distinct levels of differentiation in vitro into the rat carotid artery submitted to endothelial injury.24 25 The implanted SMCs were marked with PKH-26, a lipophilic cell membrane linker that is halved with each cell division but is not lost from the cell membrane.34 Our results show that the 2 implanted populations essentially retain for 20 days in vivo the phenotype that they specifically exhibited in vitro.. Spindle-shaped and epithelioid rat SMC populations have ...
TY - JOUR. T1 - A 310-bp minimal promoter mediates smooth muscle cell-specific expression of telokin. AU - Smith, Aiping F.. AU - Bigsby, Robert M.. AU - Word, R. Ann. AU - Herring, B. Paul. PY - 1998/5/1. Y1 - 1998/5/1. N2 - A cell-specific promoter located in an intron of the smooth muscle myosin light chain kinase gene directs transcription of telokin exclusively in smooth muscle cells. Transgenic mice were generated in which a 310-bp rabbit telokin promoter fragment, extending from -163 to +147, was used to drive expression of simian virus 40 large T antigen. Smooth muscle-specific expression of the T-antigen transgene paralleled that of the endogenous telokin gene in all smooth muscle tissues except uterus. The 310-bp promoter fragment resulted in very low levels of transgene expression in uterus; in contrast, a transgene driven by a 2.4-kb fragment (-2250 to +147) resulted in high levels of transgene expression in uterine smooth muscle. Telokin expression levels correlate with the estrogen ...
Methods and Results-Ex vivo optical imaging confirmed that Id3−/− Ldlr−/− mice have significantly fewer aortic B cells than Id3+/+ Ldlr−/− mice. After 8 and 16 weeks of Western diet, Id3−/− Ldlr−/− mice developed significantly more atherosclerosis than Id3+/+ Ldlr−/− mice, with Id3+/− Ldlr−/− mice demonstrating an intermediate phenotype. There were no differences in serum lipid levels between genotypes. Immunostaining demonstrated that aortas from Id3−/− Ldlr−/− mice had greater intimal macrophage density and C-C chemokine ligand 20 and vascular cell adhesion molecule 1 (VCAM-1) expression compared with Id3+/+ Ldlr−/− mice. Real-time polymerase chain reaction demonstrated increased VCAM-1 mRNA levels in the aortas of Id3−/− Ldlr−/− mice. Primary vascular smooth muscle cells from Id3−/− mice expressed greater amounts of VCAM-1 protein compared with control. Gain and loss of function studies in primary vascular smooth muscle cells identified a ...
Citation: Kumari, R. et al. (2003) ATP and UTP responses of cultured rat aortic smooth muscle cells revisited: Dominance of P2Y2 receptors. British Journal of Pharmacology, 140 (7), pp. 1169-1176. ...
Vascular smooth muscle cells contribute to the formation of atherosclerotic plaques by proliferating in response to vascular injury and releasing growth-promoting factors. Because their autocrine and paracrine effects are not fully understood, expression of such growth factor genes in specific cell types in vivo would help to determine their mechanism of action. We describe a method to transfer vascular smooth muscle cells expressing recombinant gene products to localized segments of the arterial wall. Vascular smooth muscle cells from the inbred Yucatan minipig were infected in vitro with an amphotropic, replication-defective retrovirus transducing the gene for Escherichia coli beta-galactosidase. Vascular smooth muscle cells expressing this recombinant gene were implanted, using a catheter, into denuded iliofemoral artery segments of pigs in vivo. These arteries subsequently demonstrated beta-galactosidase activity in cells of the intima and media. This method, which provides for the ...
Atherosclerosis is an inflammatory disease that is characterised by the involvement of chemokines that are important for the recruitment of leukocytes and scavenger receptors that mediate foam cell formation. Several cytokines are involved in the regulation of chemokines and scavenger receptors in atherosclerosis. CXCL16 is a chemokine and scavenger receptor and found in macrophages in human atherosclerotic lesions. Using double-labelled immunohistochemistry, we identified that smooth muscle cells in human lesions express CXCL16. We then analysed the effects of IFN-gamma, TNF-alpha, IL-12, IL-15, IL-18, and LPS on CXCL16 expression in cultured aortic smooth muscle cells. IFN-gamma was the most potent CXCL16 inducer and increased mRNA, soluble form, membrane form, and total cellular levels of CXCL16. The IFN-gamma induction of CXCL16 was also associated with increased uptake of oxLDL into these cells. Taken together, smooth muscle cells express CXCL16 in atherosclerotic lesions, which may play a ...
The present study examined age-related changes in the vascular relaxation response to adenine nucleotides in hypertensive and normotensive rats. Aortic ring segments from normotensive Wistar-Kyoto (WKY) rats and spontaneously hypertensive rats (SHR), age 4-6, 9-10, and 13-14 weeks, were examined for relaxation to adenosine 5-triphosphate (ATP). The extent of ATP-induced relaxation in aortic ring segments with intact endothelium was unchanged with advancing age. Rubbed (endothelium-denuded) ring preparations at the age of 4-6 weeks showed a dose-dependent relaxation similar to that of the unrubbed rings. With advancing age, the ATP-induced relaxation in the rubbed rings decreased and was abolished. The relaxation response did not differ between the SHR and WKY animals at any age, whether the preparations were rubbed or unrubbed. The stable ATP analogue beta,r-methylene ATP induced a relaxation response similar to ATP in rubbed rings at 4-6 weeks of age. In addition, treatment with ...
TY - JOUR. T1 - Modulation of collagen synthesis by tumor necrosis factor alpha in cultured vascular smooth muscle cells. AU - Hiraga, Syouichi. AU - Kaji, Toshiyuki. AU - Ueda, Yoshimichi. AU - Zisaki, Fumiko. AU - Iwata, Kazushi. AU - Koizumi, Fumitomo. AU - Okada, Yasunori. AU - Katsuda, Shogo. AU - Nakanishi, Isao. PY - 1999/12/10. Y1 - 1999/12/10. N2 - Collagen synthesis in vascular smooth muscle cells (SMCs) after exposure to tumor necrosis factor alpha (TNF-α) was investigated using a culture system. The synthesis of collagenase-digestible proteins (CDP) and noncollagenous proteins (NCP) was evaluated by the [3H]proline incorporation. It was shown that TNF-α markedly suppresses the incorporation of [3H]proline into both CDP and NCP in confluent cultures of SMCs but not in sparse cultures of the cells. Such a marked suppression by TNF-α was not observed in confluent bovine aortic endothelial cells and human fibroblastic IMR-90 cells. In confluent SMCs, the synthesis of CDP was more ...
Vascular smooth muscle (VSM) cell proliferation contributes to the pathogenesis of atherosclerosis, restenosis after angioplasty and vein graft disease. The regulation of genes involved in VSM cell proliferation, particularly by naturally occurring inhibitors, is therefore of some importance. We have investigated the role of the c-myc proto-oncogene in growth arrest of exponentially proliferating rat VSM cells, following mitogen withdrawal, treatment with heparin (50 micrograms/ml), interferon-gamma (IFN-gamma) (100 i.u./ml), or the cyclic nucleotide analogues, 8-bromo-adenosine-3′5′-cyclic monophosphate (8-Br-cAMP; 0.1 mM) and 8-bromoguanosine-3′5′-cyclic monophosphate (8-Br-cGMP; 0.1 mM). Growth arrest was accompanied by down-regulation of c-Myc protein and mRNA following treatment with all inhibitors. Serum withdrawal or IFN-gamma treatment suppressed c-myc expression by more than 50% within 2 h, and this occurred throughout the cell cycle. Platelet-derived growth factor, epidermal ...
I am planning to set up a co-culture system for human endothelial cells and human VASCULAR smooth muscle cells. I would be really interested to find out about methods of extraction and primary culture of human vascular smooth muscle cells, if anyone can help and advise, please mail me! Thanks Pippa Deex ...
Vascular smooth muscle refers to the particular type of smooth muscle found within, and composing the majority of the wall of blood vessels. Vascular smooth muscle refers to the particular type of smooth muscle found within, and composing the majority of the wall of blood vessels. Vascular smooth muscle is innervated primarily by the sympathetic nervous system through adrenergic receptors (adrenoceptors). The three types of adrenoceptors present are: α 1 {\displaystyle \alpha _{1}} , α 2 {\displaystyle \alpha _{2}} and β 2 {\displaystyle \beta _{2}} . The main endogenous agonist of these cell receptors is norepinephrine (NE). The adrenergic receptors exert opposite physiologic effects in the vascular smooth muscle under activation: α 1 {\displaystyle \alpha _{1}} receptors. Under NE binding α 1 {\displaystyle \alpha _{1}} receptors cause vasoconstriction (i.e. contraction of the vascular smooth muscle cells decreasing the diameter of the vessels). α 1 {\displaystyle \alpha _{1}} receptors ...
292653345 - EP 1085880 A2 2001-03-28 - USE OF ALKYLATING COMPOUNDS FOR INHIBITING PROLIFERATION OF ARTERIAL SMOOTH MUSCLE CELLS - [origin: WO9963981A2] The present invention provides methods and compositions for inhibiting the proliferation of smooth muscle cells at a site of vascular injury. The methods include intravascular administration of a reactive compound to the site of injury, without the requirement for activation or sustained release of the compound.[origin: WO9963981A2] The present invention provides methods and compositions for inhibiting the proliferation of smooth muscle cells at a site of vascular injury. The methods include intravascular administration of a reactive compound to the site of injury, without the requirement for activation or sustained release of the compound.
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Diabetic Mouse Brain Vascular Smooth Muscle Cells from Creative Bioarray are isolated from the brain vessel of Diabetic (db/db) mice (8 weeks). Diabetic Mouse Brain Vascular Smooth Muscle Cells are grown in T25 tissue culture flasks pre-coated with gelatin-based coating solution for 2 min and incubated in Creative Bioarrays Culture Complete Growth Medium generally for 3-7 days. Prior to shipping, cells at passage 1 are detached from the culture flasks and immediately cryo-preserved in vials. Each vial contains at least 0.5x10^6cells per ml and is delivered frozen ...
BioAssay record AID 421037 submitted by ChEMBL: Inhibition of PDGF-BB-stimulated Rac1 activity in human aortic smooth muscle cells assessed as reduction of ratio of Rac1GTP/Rac1 levels at 25 uM after 4 hrs by pull-down assay.
The present study may have important pathological and therapeutic implications because overgrowth of VSMCs is a pivotal etiologic factor in the development of atherosclerosis and restenosis after angioplasty.26-28 To date, inhibiting VSMC proliferation is among the most effective strategies for preventing their overgrowth and controlling neointimal thickening.14 Previous studies have shown that targeting Ras with negative regulators or blocking the Ras downstream pathways is able to effectively attenuate restenosis from balloon catheterization.14,15,29-33 Our recent studies have demonstrated that rMfn-2 is a powerful endogenous Ras inhibitor and that somatic gene transfer of rMfn-2 profoundly inhibits rat VSMC proliferation and balloon injury-induced neointima thickening in vivo by inhibiting the Ras-Raf-MEK-ERK/MAPK signaling pathway.17. In addition to inhibition of cell proliferation, growing evidence has indicated that apoptosis also plays an essential role in the control of neointimal ...
Aortic calcification was demonstrated in experimental animal models of hyperhomocysteinemia. Mild hyperhomocysteinemia was associated with aortic calcification, suggesting a relationship between homocysteine (HCY) and the pathogenesis of aortic calcification. In the present study, the effect of HCY on vascular calcification was examined in calcifying and non-calcifying vascular smooth muscle cells (VSMCs). Cell calcification was induced by incubation of VSMCs with [ glycerophosphate. Proliferation of VSMCs was studied by cell counting, H-3-thymidine (H-3-TdR) and H-3-leucine (H-3-Leu) incorporation. Ca-45 accumulation, cell calcium content, and alkaline phosphatase (ALP) activity were measured as indices of calcification. The results showed that the proliferation of calcifying VSMCs, which was indicated by cell counting, H-3-TdR and H-3-Leu incorporation in calcifying VSMCs, was enhanced as compared with that of non-calcifying VSMCs. HCY promoted increases in cell number, H-3-TdR and H-3-Leu ...
Diabetic complications largely affect the circulation and are associated with resistance to insulin and altered levels of insulin-like growth factor-I (IGF-I). Insulin resistance and altered IGF-I levels are also associated with vascular disease. Insulin and IGF-I are highly homologous peptides and can cross react with each others respective receptors, insulin receptors (IR) and IGF-I receptors (IGFIR), which also share homology to a large extent and can form hybrid IR/IGF-IR. Cultured endothelial and vascular smooth muscle cells from different vascular beds express considerably more IGF-IR than IR. Since the direct action of insulin and IGFs on the vasculature remains poorly understood, our aim was to study mechanisms behind insulin resistance and IGF-I sensitivity and the possible impact of hybrid IR/IGF-IR in vascular cells.. This thesis is based on four papers investigating the presence of IR and IGF-IR in cultured endothelial and vascular smooth muscle cells, and in tissue specimens from ...
TY - GEN. T1 - Curvature-induced spontaneous detachment of vascular smooth muscle cell sheets. T2 - Towards vascular self assembly in microchannels. AU - Yamashita, Tadahiro. AU - Kollmannsberger, P.. AU - Mawatari, K.. AU - Vogel, V.. AU - Kitamori, T.. PY - 2013/1/1. Y1 - 2013/1/1. N2 - A new model is proposed which describes the spontaneous detachment of vascular smooth muscle cells induced by surface curvature. Growing tubular structures from smooth muscle cells (SMCs) in vitro is a key challenge in microvascular tissue engineering. SMC growth is however significantly suppressed on curved substrates. We show that this is caused by mechanical interaction between adhering cells and the surrounding geometry, which compromises the adhesion of growing tissue. Our model opens up new strategies for engineering luminal vasculature in microdevices, and gives new insights for controlling tissue formation in micro environments.. AB - A new model is proposed which describes the spontaneous detachment of ...
BACKGROUND: Pathological vascular remodeling in venous bypass grafts (VGs) results in smooth muscle cell (SMC) intimal hyperplasia and provides the substrate for progressive atherosclerosis, the principal cause of late VG failure. Nitric oxide (NO) bioactivity is reduced in VGs, in association with increased vascular superoxide production, but how these features relate to pathological VG remodeling remains unclear. We used gene transfer of the neuronal isoform of nitric oxide synthase (nNOS) to investigate how increased NO production modulates vascular remodeling in VGs and determined the effects on late VG phenotype. METHODS AND RESULTS: New Zealand White rabbits (n=60) underwent jugular-carotid interposition bypass graft surgery with intraoperative adenoviral gene transfer of nNOS or beta-galactosidase. Vessels were analyzed after 3 days (early, to investigate acute injury/inflammation) or 28 days (late, to investigate SMC intimal hyperplasia). In early VGs, nNOS gene transfer significantly increased
MicroRNAs (miRNAs) are an emerging class of highly conserved, non-coding small RNAs that regulate gene expression at the post-transcriptional level. Recent findings have shown that miR-1 and miR-133 play a critical role in cardiogenesis and cardiomyocyte hypertrophy. However, the role of miR-1 and miR-133 in vascular disease is currently unknown. Thus, the aim of the present study was to evaluate the role, if any, of miR-1 and miR-133 in vascular smooth muscle cell (VSMC) growth in vitro and in vivo. miR-1 and miR-133 transcripts were evaluated by quantitative real-time polymerase chain reaction (qRT-PCR) in quiescent vs. proliferating VSMCs in vitro and in vivo. VSMC were transfected in culture dishes with adenoviral vector constructs carrying miR-1 or miR133. VSMC proliferation was measured by Brdu incorporation. VSMC apoptosis was induced by H2O2 and measured by a Tdt assay. In the in vivo protocol, balloon injury of the right carotid was produced in male Wistar rats. Straight after the ...
Little, Peter J., Ballinger, Mandy L., Survase, Soniya, Osman, Narin, Ogru, Esra, Geytenbeek, Stephen, Bruemmer, Dennis and Nigro, Julie (2008) Phosphorylated troglitazone activates PPAR gamma and inhibits vascular smooth muscle cell proliferation and proteoglycan synthesis. Journal of Cardiovascular Pharmacology, 51 3: 274-279. ...
Objective: Peroxisome proliferator-activated receptor γ (PPARγ) agonists reduce blood pressure (BP) and vascular injury in hypertensive rodents and humans. Pparγ inactivation in vascular smooth muscle cells (VSMC)using a tamoxifen inducible Cre-Lox system enhanced angiotensin II-induced vascular injury. Transgenic mice overexpressing endothelin (ET)-1selectively in the endothelium (eET-1) exhibit endothelial dysfunction, increased oxidative stress and inflammation. We hypothesized that inactivation of Pparγ in VSMC(smPparγ-/-)will exaggerate ET-1-induced vascular damage.. Methods and Results: Elevenweek-old male control, eET-1, smPparγ-/-and eET-1/smPparγ-/- mice weretreated with tamoxifen (1 mg/kg/day, s.c.) for 5 days and sacrificed 4 weeks later. Systolic BP was higher in eET-1compared to control (123±5 vs 109±2 mmHg,P,0.05)and unaffected by Pparγ inactivation.Mesenteric artery (MA) vasodilatory responses to acetylcholine were impaired only in smPparγ-/- (P,0.05) compared to ...
The recognition that cells of the vascular wall can secrete cytokines such as IL-1 suggests new mechanisms for initiating or sustaining inflammatory responses in blood vessels. We report that purified human monocyte-derived IL-1 or recombinant human IL-1 (rIL-1 beta and rIL-1 alpha) induce cultured human smooth muscle cells derived from veins or arteries to synthesize IL-1 beta mRNA and produce and release biologically active IL-1. rIL-1 beta also stimulated the production of PGE2 by smooth muscle cells. Exposure to rIL-1 beta (1-100 ng/ml), or rIL-1 alpha (0.01-10 ng/ml) increased IL-1 beta mRNA levels within 30 min. Actinomycin D (1 microgram/ml) prevented the induction of IL-1 beta mRNA by rIL-1. IL-1 alpha mRNA was detected in SMC treated with cycloheximide (1 microgram/ml) and rIL-1 beta, or cycloheximide alone. rIL-1 alpha and rIL-1 beta produced maximal levels of IL-1 beta mRNA after 4 h, and intracellular IL-1 biological activity after 6 h of exposure. Release of IL-1 activity in the ...
OBJECTIVE: Cerebral aneurysm is a common vascular disease with high morbidity and mortality. Vascular smooth muscle deletion or dysplasia is an important r
Methods Firstly, we detected the expression of lincRNA-p21 expression in the aortic plaque of apoE-/- mice fed with high-fat diet and peripheral blood mononuclear cells of clinical coronary disease patients. Then we observed the role of lincRNA-p21 in cell proliferation and apoptosis using mice macrophage cell line RAW264.7 and human vascular smooth muscle cell line HA-VSMCs by loss-of-function and gain-of function approaches. Meanwhile, the expression of mRNA and protein levels of apoptosis-related downstream targeting of p53 also been detected. Furthermore, we preformed bioinformatics prediction, RNA-Immunoprecipitation (RIP), RNA-pulldown and deletion mapping experiments to test the potential interaction and specific interaction pattern between lincRNA-p21 and MDM2. Then co-Immunoprecipitaiton (co-IP) and Chromatin- Immunoprecipitation (ChIP) assays were preformed to verify the possible influence on p53 transcriptional activity of this binding. We further investigated whether or not ...
Maspin is a serpin that has multiple effects on cell behavior, including inhibition of migration. How maspin mediates these diverse effects remains unclear, as it is devoid of protease inhibitory activity. We have previously shown that maspin rapidly inhibits the migration of vascular smooth muscle cells (VSMC), suggesting the involvement of direct interactions with cell surface proteins. Here, using immunofluorescence microscopy, we demonstrate that maspin binds specifically to the surface of VSMC in the dedifferentiated, but not the differentiated, phenotype. Ligand blotting of VSMC lysates revealed the presence of several maspin-binding proteins, with a protein of 150 kDa differentially expressed between the two VSMC phenotypes. Western blotting suggested that this protein was the beta1 integrin subunit, and subsequently both alpha3beta1 and alpha5beta1, but not alphavbeta3, were shown to associate with maspin by coimmunoprecipitation. Specific binding of these integrins was also observed ...
Ca2+ permeability of the alkalosis-induced membrane conductance was clearly evident from an approximately 80-mV negative shift in reversal potentials associated with a reduction of extracellular Ca2+ by two orders of magnitude (0 Na+/2 Ca2+ versus 0 Na+/0.02 Ca2+ solution). The cation channels underlying this conductance were barely voltage-dependent and insensitive to the classical Ca2+ channel blocker verapamil but sensitive to inhibition by 2-APB, which is known to interfere with IP3 receptor function22 and to inhibit store-dependent Ca2+ entry pathways.23 NH4Cl-induced, 2-APB-sensitive Ca2+ entry was also observed in primary rat aortic cells (online data supplement), and is therefore unlikely a phenomenon specific for the A7r5 cell line. Effects of 2-APB may be mediated by a direct block of the Ca2+ entry channels as recently suggested for capacitative Ca2+ entry.23 Interestingly, NH4Cl-induced intracellular Ca2+ mobilization in Ca2+ free solution was not significantly inhibited by 2-APB ...
The relationship of Na+K+ATPase and hypertension has been a topic of concentrated study for the past twenty years. During the onset of hypertension, an increase in Na+K+ATPase in the cellular membranes of the aortic smooth muscle cells has been found to occur. The reason for this increased manifestation of Na+K+ATPase during the beginning stages of hypertension and how the cells regulate this mechanism are still unclear. An initial stage of hypertension is the increase in blood pressure, which contracts the aortic smooth muscle cell lining. Previous in vitro studies have shown the contraction of the aortic smooth muscle cells does demonstrate this increased expression of Na+K+ATPase. The objective of this study was to begin to understand the regulation of the increased expression by beginning with mRNAs control and regulation. Using a Flexercell Strain Unit which cyclically stretches cells, the mRNA expression during 20%, 10%, and non-stretch controls were measured by analysis of Northern ...
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The effect of simvastatin, atrovastatin and pravastatin as well as of the proteasom inhibitor beta-clasto lactacystin was studied on morphology, proliferation viability and on proteasomaö activityin two mammalian endothelial cell lines (CPAE and Ey.hy926) and in primary vascular smooth muscle cells (VSMCs). Both statins and lactacystin induced comparable morphological changes and attenuated proliferation of CPAE. Whereas the statin-induced effects were reversed by mevalonic acid, however, the lactacystin-induced alterations were not influenced by mevalonic acid. As expected, lactacystin caused a significant loss of proteasomal activity measured in the extracts of trested cells. The extracts of statin-treated CPAEs exhibited unchanged activities. This result was also confirmed in Ea.hy926 cells and in primary rat VSMCs. It is shown, that even high dosis of statins do not modulate the activity of purified human 20S proteasomes. The conclusion was that similar biological effects of statins and the ...
We previously reported a relationship between forearm resistance vessel function and global neuropsychological performance in patients with atherosclerotic vascular disease (AVD). This study was conducted to determine the relationships among vascular smooth muscle function, endothelial function, and initiation and processing speed in this sample. Participants were 80 individuals with AVD. Resistance vessel function was measured before and after infusion of vasoactive agents. Neuropsychological assessment included measures of estimated premorbid cognitive function, current global cognitive function, initiation, and processing speed. Vascular smooth muscle function was significantly associated with the initiation/processing speed composite score [R-Square Change = .152; F Change (1,71) = 16.61; p , .001], above and beyond the variance accounted for by age, education, premorbid cognitive function, and endothelium-dependent vascular function. This relationship remained significant when controlling ...
The vascular smooth muscle cell (VSMC) is a highly specialized cell whose principal function is contraction. On contraction, VSMCs shorten, thereby decreasing the diameter of a blood vessel to regulate the blood flow and pressure. The principal mechanisms that regulate the contractile state of VSMCs are changes in cytosolic Ca2+ concentration ([Ca2+]c). In response to vasoconstrictor stimuli, Ca2+ is mobilized from intracellular stores and/or the extracellular space to increase [Ca2+]c in VSMCs. The increase in [Ca2+]c, in turn, activates the Ca2+-CaM-MLCK pathway and stimulates MLC20 phosphorylation, leading to myosin-actin interactions and, hence, the development of contractile force. The sensitivity of contractile myofilaments or MLC20 phosphorylation to Ca2+ can be secondarily modulated by other signaling pathways. During receptor stimulation, the contractile force is greatly enhanced by the inhibition of myosin phosphatase. Rho/Rho kinase, PKC, and arachidonic acid have been proposed to ...
Vascular smooth muscle contraction is mediated by activation of extracellular signal-regulated kinase (ERK) 1/2, an isoform of mitogen-activated protein kinase (MAPK). However, the role of stress-activated protein kinase/,i,c-Jun,/i, N-terminal kinase (JNK) in vascular smooth muscle contraction has not been defined. We investigated the role of JNK in the contractile response to norepinephrine (NE) in rat aortic smooth muscle. NE evoked contraction in a dose-dependent manner, and this effect was inhibited by the JNK inhibitor SP600125. NE increased the phosphorylation of JNK, which was greater in aortic smooth muscle from hypertensive rats than from normotensive rats. NE-induced JNK phosphorylation was significantly inhibited by SP600125 and the conventional-type PKC (cPKC) inhibitor Gö6976, but not by the Rho kinase inhibitor Y27632 or the phosphatidylinositol 3-kinase inhibitor LY294002. Thymeleatoxin, a selective activator of cPKC, increased JNK phosphorylation, which was inhibited by ...
Lawson, J S and Syme, H M and Wheeler-Jones, C P D and Elliott, J (2019) Characterisation of Crandell-Rees Feline Kidney (CRFK) cells as mesenchymal in phenotype. RESEARCH IN VETERINARY SCIENCE, 127. pp. 99-102. Patel, J J and Bourne, L E and Millán, J L and Arnett, T R and MacRae, V E and Wheeler-Jones, C P D and Orriss, I R (2019) Inhibition of vascular smooth muscle cell calcification by ATP-analogues. Purinergic Signalling, 15 (3). pp. 315-326. Patel, J J and Bourne, L E and Davies, B K and Arnett, T R and MacRae, V E and Wheeler-Jones, C P D and Orriss, I R (2019) Differing calcification processes in cultured vascular smooth muscle cells and osteoblasts. Experimental Cell Research, 380 (1). pp. 100-113. Lawson, J S and Liu, H-H and Syme, H M and Purcell, R and Wheeler-Jones, C P D and Elliott, J (2018) The cat as a naturally occurring model of renal interstitial fibrosis: Characterisation of primary feline proximal tubular epithelial cells and comparative pro-fibrotic effects of TGF-ß1. ...
Apoptosis of vascular smooth muscle cells induces features of plaque vulnerability in atherosclerosis. Connexin37 protects against atherosclerosis by regulating monocyte adhesion
Fibronectin-splice variant containing extra domain A (Fn-EDA) is associated with smooth muscle cells (SMCs) following vascular injury. The role of SMC-derived Fn-EDA in SMC phenotypic switching or its implication in neointimal hyperplasia remains unclear. Herein, using human coronary artery sections with a bare metal stent, we demonstrate the expression of Fn-EDA in the vicinity of SMC-rich neointima and peri-strut areas. In mice, Fn-EDA colocalizes with SMCs in the neointima of injured carotid arteries and promotes neointima formation in the comorbid condition of hyperlipidemia by potentiating SMC proliferation and migration. No sex-based differences were observed. Mechanistic studies suggested that Fn-EDA mediates integrin- and TLR4-dependent proliferation and migration through activation of FAK/Src and Akt1/mTOR signaling, respectively. Specific deletion of Fn-EDA in SMCs, but not in endothelial cells, reduced intimal hyperplasia and suppressed the SMC synthetic phenotype concomitant with ...
Fibronectin-splice variant containing extra domain A (Fn-EDA) is associated with smooth muscle cells (SMCs) following vascular injury. The role of SMC-derived Fn-EDA in SMC phenotypic switching or its implication in neointimal hyperplasia remains unclear. Herein, using human coronary artery sections with a bare metal stent, we demonstrate the expression of Fn-EDA in the vicinity of SMC-rich neointima and peri-strut areas. In mice, Fn-EDA colocalizes with SMCs in the neointima of injured carotid arteries and promotes neointima formation in the comorbid condition of hyperlipidemia by potentiating SMC proliferation and migration. No sex-based differences were observed. Mechanistic studies suggested that Fn-EDA mediates integrin- and TLR4-dependent proliferation and migration through activation of FAK/Src and Akt1/mTOR signaling, respectively. Specific deletion of Fn-EDA in SMCs, but not in endothelial cells, reduced intimal hyperplasia and suppressed the SMC synthetic phenotype concomitant with ...
by Huei-Ping Tzeng, Kuo-Cheng Lan, Ting-Hua Yang, Min-Ni Chung, Shing Hwa Liu Benzo[a]pyrene, a ubiquitous environmental pollutant, has been suggested to be capable of initiating and/or accelerating atherosclerosis. Accumulation of vascular smooth muscle cells (VSMCs) in vessel intima is a hallmark of atherosclerosis. Nitric oxide (NO) can suppress VSMCs proliferation and induce VSMCs apoptosis. NO…
TY - JOUR. T1 - Calcium sparklets regulate local and global calcium in murine arterial smooth muscle. AU - Amberg, Gregory C.. AU - Navedo, Manuel F.. AU - Nieves-cintrón, Madeline. AU - Molkentin, Jeffery D.. AU - Santana, Luis F.. PY - 2007/2/15. Y1 - 2007/2/15. N2 - In arterial smooth muscle, protein kinase Cα (PKCα) coerces discrete clusters of L-type Ca2+ channels to operate in a high open probability mode, resulting in subcellular domains of nearly continual Ca2+ influx called persistent Ca2+ sparklets. Our previous work suggested that steady-state Ca2+ entry into arterial myocytes, and thus global [Ca2+]i, is regulated by Ca2+ influx through clusters of L-type Ca2+ channels operating in this persistently active mode in addition to openings of solitary channels functioning in a low-activity mode. Here, we provide the first direct evidence supporting this Ca2+ sparklet model of Ca2+ influx at a physiological membrane potential and external Ca2+ concentration. In support of this ...
In this study, we demonstrated that Bo-Gan-Whan (BGH), a Korean polyherbal medicine, has an inhibitory effect on VSMC migration and proliferation in response to PDGF-BB as revealed by the results obtained from the scratch-wound healing and Boyden chamber assay and sprout aortic ring assays, respectively. Moreover, it was demonstrated through western blot analysis that the modulation of MAPKs is the major signal that is activated in the pathogenesis of VSMCs through activation of the ERK1/2 and p38 MAPK pathways. These results were confirmed from the ex vivo analysis of PDGF-BB-induced VSMCs migration and proliferation. The data on ex vivo analysis, through outgrowth of vessel sprouts from the aortic strips assay, show that BGH treatment can significantly reduce VSMC migration and proliferation after PDGF-BB stimulation.. Abnormal proliferation of VSMCs is a key to the vascular pathological conditions such as atherosclerosis and restenosis. Moreover, excessive migration of VSMCs in vascular ...
Background MEK1/2 is a serine/threonine protein that phosphorylates extracellular signal-regulated kinase (ERK1/2). Cerebral ischemia results in enhanced expression of cerebrovascular contractile...
The Microcirculatory Core Laboratorys primary mission is to study regulation of cardiovascular resistance at the vascular level. Our experties contribute to the mission of the Consortium for Integrative Cardiovascular Research by providing a intermediate level of investigation between integrative (intact animal and whole organ) and vascular biology (isolated endothelial and vascular smooth muscle cell) studies.. Research Interests The Microcirculatory Core Laboratory specialises in isolated vascular preparations to study intrinsic regulation of vascular tone. We are interested in endothelial and smooth muscle cell function in health and disease. We predominantly study resistance vessels (arterioles and small arteries), since these are primary determinants of resistance in the cardiovascular circuit. We use disease models where resistance vascular function is altered, such as experimental models of hemorrhagic shock, hypertension, obesity, insulin resistance, and diabetes. ...
Nitroalkenes, the nitration products of unsaturated fatty acids formed via NO-dependent oxidative reactions, have been demonstrated to exert strong biological actions in endothelial cells and monocytes/macrophages; however, little is known about thei