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1.
The localization of neuropeptide Y (NPY) and atrial natriuretic peptide (ANP) in the endothelial cells of human umbilical blood vessels was studied using the pre-embedding peroxidase-antiperoxidase (PAP) technique for electron microscopy and avidin-biotin-complex (ABC) immunostaining for endothelial cells cultured from umbilical vein. Subpopulations of NPY- and ANP-immunoreactive endothelial cells were present in term umbilical vein and artery. The umbilical vein contained more positive cells than the artery. The percentage of NPY- and ANP-immunoreactive umbilical vein cells in culture was 32% and 44%, respectively, out of a total of 3013 cells examined. The possibility that these potent vasoactive substances located in the endothelial cells of the non-innervated umbilical vessels are involved in the local regulation of blood flow is discussed. 相似文献
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W. Q. Cai K. Dikranian P. Bodin M. Turmaine G. Burnstock 《Cell and tissue research》1993,274(3):533-538
Human umbilical vessels are unique in lacking any innervation; thus endothelial cells may play the major role in local control and regulation of the blood flow. In the present study, we examined ultrathin sections of cultured human umbilical vein endothelial cells and tissue preparations of umbilical vein and artery, immunostained by the post-embedding colloidal gold double-labelling technique. We observed colocalization of atrial natriuretic peptide and neuropeptide Y, as well as colocalization of atrial natriuretic peptide and neuropeptide Y with other vasoactive substances, namely, vasoactive intestinal peptide, substance P, calcitonin gene-related peptide and arginine vasopressin. The functional significance of the colocalization of these vasoactive substances in the human umbilical vessel endothelial cells is discussed. 相似文献
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Vasoactive peptides upregulate mRNA expression and secretion of vascular endothelial growth factor in human airway smooth muscle cells 总被引:2,自引:0,他引:2
Alagappan VK Willems-Widyastuti A Seynhaeve AL Garrelds IM ten Hagen TL Saxena PR Sharma HS 《Cell biochemistry and biophysics》2007,47(1):109-118
Airway remodeling and associated angiogenesis are documented features of asthma, of which the molecular mechanisms are not
fully understood. Angiotensin (ANG)II and endothelin (ET)-1 are potent vasoconstricting circulatory hormones implicated in
asthma. We investigated the effects of ANG II and ET-1 on human airway smooth muscle (ASM) cells proliferation and growth
and examined the mRNA expression and release of the angiogenic peptide, vascular endothelial growth factor (VEGF). Serum deprived
(48 h) human ASM cells were incubated with ANG II (100 nM) or ET-1 (10nM) for 30 min, 1, 2, 4, 8, 16, and 24 h and the endogenous synthesis of VEGF was examined in relation to control cells receiving
serum free culture medium. ET-1 induced time dependent DNA biosynthesis as determined by [3H]-thymidine incorporation assay. Using northern blot hybridization, we detected two mRNA species of 3.9 and 1.7 kb encoding
VEGF in the cultured smooth muscle cells. Both ANG II and ET-1 induced the mRNA expression (two-to threefold) and secretion
(1.8-to 2.8-fold) of VEGF reaching maximal levels between 4–8 h of incubation. Induced expression and release of VEGF declined
after 8 h of ANG II incubation while levels remained elevated in the case of ET-1. The conditioned medium derived from ET-1-treated
ASM cells induced [3H]-thymidine incorporation and cell number in porcine pulmonary artery endothelial as well as human umbilical vein endothelial
cells. Moreover, the VEGF tyrosine kinase receptor inhibitor blocked the conditioned medium induced mitogenesis in endothelial
cells. Our results suggest a potential role for ANG II and ET-1 in ASM cell growth and upregulation of VEGF that may participate
in endothelial cell proliferation via paracrine mechanisms and thus causing pathological angiogenesis and vascular remodelling
seen during asthma. 相似文献
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Expression of serotonin receptor mRNAs in blood vessels 总被引:17,自引:0,他引:17
Using RT-PCR we distinguished mRNAs for all known G-protein coupled serotonin receptors expressed in various rat and porcine blood vessels. Nearly all vessels expressed 5HT1
β, 5-HT2A, 5-HT2B, 5-HT4, and 5-Ht7 receptor mRNA to different extents. New splice variants of the porcine 5-HT4 receptor were observed. Similar PCR assays were performed with endothelial and smooth muscle cells from human pulmonary artery, aorta, and with endothelial cells from human coronary artery and umbilical vein. All endothelial cells expressed 5-HT1
β, 5-HT2b, and 5-HT4 receptor mRNA, whereas in smooth muscle cells 5-HT1
β, 5-HT2A, 5-HT7, and in some experiments 5-HT2B receptor mRNA were found. A model for the regulation of vascular tone by different 5-HT receptors is proposed. 相似文献
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Kook H Itoh H Choi BS Sawada N Doi K Hwang TJ Kim KK Arai H Baik YH Nakao K 《American journal of physiology. Heart and circulatory physiology》2003,284(4):H1388-H1397
Both nitric oxide (NO) and natriuretic peptides produce apoptosis of vascular smooth muscle cells. However, there is evidence that NO induces endothelial cell proliferation, which suggests that there is a difference in the response of endothelial cells to natriuretic peptides. The purpose of this study was to investigate the effect of atrial natriuretic peptide (ANP) on human endothelial cell survival. ANP within the physiological concentration (10(-11) mol/l) induced a 52% increase in the number of human coronary arterial endothelial cells and a 63% increase in human umbilical vein endothelial cells at a low concentration of serum. The increase in cell numbers was blocked by pretreatment with RP8-CPT-cGMP (RP8), a cGMP-dependent protein kinase inhibitor, with wortmannin, an Akt/PKB inhibitor, and with PD-98059, an ERK1/2 inhibitor. In a Transwell migration test, ANP also increased the cell migration, and RP8, wortmannin, and PD-98059 blocked this increase. A wound healing assay was performed to examine the effects of ANP on regeneration in vitro. ANP increased both cell numbers and migration, but the effects were blocked by the above three kinase inhibitors. ANP increased the expression of phospho-Akt and of phospho-ERK1/2 within 1.5 h. These results suggest that ANP can potentiate endothelial regeneration by cGMP-dependent protein kinase stimulation and subsequent Akt and ERK1/2 activations. 相似文献
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Expression of TRPC homologs in endothelial cells and smooth muscle layers of human arteries 总被引:1,自引:1,他引:0
Yip H Chan WY Leung PC Kwan HY Liu C Huang Y Michel V Yew DT Yao X 《Histochemistry and cell biology》2004,122(6):553-561
TRPC channels are a group of Ca2+-permeable nonselective cation channels that mediate store-operated and/or agonist-stimulated Ca2+ influx in a variety of cell types. In this study, we extensively examined the expression patterns of TRPC homologs in human vascular tissues. RT-PCR amplified cDNA fragments of TRPC1 (505 bp), TRPC3 (372 bp), TRPC4 (499 bp), TRPC5 (325 bp), TRPC6 (509 bp), and TRPC7 (187 bp) from RNA isolated from cultured human coronary artery endothelial cells. In situ hybridization yielded strong labeling of TRPC1,3–6 in the endothelial and smooth muscle cells of human coronary and cerebral arteries. TRPC7 labeling was exclusively found in endothelial cells but not in smooth muscle cells. Results from immunohistochemical staining were consistent with those from in situ hybridization. Similar expression patterns of TRPC homologs were also observed in arterioles and vaso vasora. In conclusion, our study indicates that TRPC homologs are widely expressed in human vessels of all calibers, including medium-sized coronary arteries and cerebral arteries, smaller-sized resistance arteries, and vaso vasora. These results suggest a ubiquitous role of TRPC homologs in regulating blood supply to different regions and in controlling arterial blood pressure. 相似文献
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A. J. Sexton A. Loesch M. Turmaine S. Miah G. Burnstock 《Cell and tissue research》1996,284(1):167-175
Human umbilical vessels are devoid of nerves and therefore endothelial cells may play an important role in the control of
feto-placental blood flow. The pharmacological effects of 5-hydroxytryptamine, histamine and endothelin were examined in umbilical
arteries and veins from legal terminations (gestational age 8–17 weeks, n=12) and normal term vaginal deliveries (gestational age 38–41, n=12). Immunocytochemistry of human unbilical vessels indicated that 5-hydroxytryptamine, histamine and endothelin were localised
in subpopulations of endothelial cells of both artery and vein in late, but not early, pregnancy. 5-Hydroxytryptamine (10
nM–30 μM) caused sustained concentration-dependent contractions in all vessels from early and late pregnancy. Histamine (0.1
μM–30 mM) also caused sustained contractions in all vessels from late pregnancy but only 27% of arteries and 41% of veins
from early pregnancy responded. Endothelin (10 pM–30 nM) caused slow long-lasting contractions in all vessels from early and
late pregnancy. Atrial natriuretic peptide and neuropeptide Y did not alter vascular tone. The endothelium may thus play an
autocrine/paracrine role, by synthesizing and releasing the above reactive substances in late pregnancy to influence feto-placental
blood flow.
Received: 23 May 1995 / Accepted: 13 October 1995 相似文献
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缺氧条件下血管活性因子对血管内皮细胞中VEGF表达的影响 总被引:4,自引:0,他引:4
探讨在缺氧条件下人脐静脉血管内皮细胞对血管内皮生长因子 (vascular endothelialgrowth factor,VEGF)表达及缩血管活性物质内皮素 (ET)、舒血管活性物质一氧化氮 (NO)和 NO抑制剂 LNNA对 VEGF基因表达的影响 .体外培养人脐静脉血管内皮细胞 ,经缺氧及血管活性物质处理 .Northern杂交、酶联免疫检测和计算机图象分析等观察 VEGF m RNA和蛋白表达水平 .发现缺氧 6h内皮细胞可见 VEGF表达 .ET可促进 VEGF m RNA的表达 ,NO可明显抑制 VEGFm RNA的表达 ,NO抑制剂 LNNA也影响 VEGF m RNA的表达 .ELISA检测 VEGF蛋白水平分别为 6h组 8.2± 1 .1 ng/ L,ET+6h组 9.37± 1 .0 2 ng/ L,NO+6h组 2 .86± 0 .91 ng/ L,L - NNA+6h组 1 4.75± 1 .87ng/ L.缺氧可诱导人脐静脉血管内皮细胞分泌 VEGF并受血管活性物质ET和 NO的调控 ,ET促进其表达 ,NO抑制其表达 . 相似文献
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Nathan C. Lo Nancy A. Turner Miguel A. Cruz Joel Moake 《The Journal of biological chemistry》2013,288(46):33118-33123
Shiga toxin (Stx) produced by enterohemorrhagic Escherichia coli causes diarrhea-associated hemolytic-uremic syndrome (DHUS), a severe renal thrombotic microangiopathy. We investigated the interaction between Stx and von Willebrand Factor (VWF), a multimeric plasma glycoprotein that mediates platelet adhesion, activation, and aggregation. Stx bound to ultra-large VWF (ULVWF) secreted from and anchored to stimulated human umbilical vein endothelial cells, as well as to immobilized VWF-rich human umbilical vein endothelial cell supernatant. This Stx binding was localized to the A1 and A2 domain of VWF monomeric subunits and reduced the rate of ADAMTS-13-mediated cleavage of the Tyr1605-Met1606 peptide bond in the A2 domain. Stx-VWF interaction and the associated delay in ADAMTS-13-mediated cleavage of VWF may contribute to the pathophysiology of DHUS. 相似文献
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K Kaji 《Human cell》1988,1(2):188-197
The purpose of this review is to introduce a simple and inexpensive culture method for human umbilical blood vessel endothelial cells. The medium used is MCDB-104 supplemented with 10% fetal bovine serum, 70 ng/ml endothelial cell growth factor from new-born bovine brains, 10 ng/ml murine epidermal growth factor, and 100 micrograms/ml heparin. The culture dishes are coated with gelatin. Under these conditions, endothelial cells from human vessels were grown with doubling times of 18-22 hrs and reached saturation densities of 8-12 x 10(4) cells/cm2. To determine the lifespan of the endothelial cells, the cells were serially subcultivated weekly at an inoculum size of 1,000 cells/cm2. Human endothelial cells from umbilical vein and artery were grown for 21 to 37 passages with 55 to 125 population doublings. This culture method seems to be useful for studying cell proliferation and functions of human endothelial cells. 相似文献
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目的探讨天然和氧化低密度脂蛋白(n-LDL,ox-LDL)对人脐静脉内皮细胞(HUVEC)VCAM-1表达的影响。方法将n-LDL,ox-LDL作用于培养的HUVEC,用细胞酶联免疫吸附试验(cellELISA)检测VCAM-1蛋白的表达,用原位分子杂交技术检测VCAM-1 mRNA的表达。结果正常培养的HUVEC可表达VCAM-1,n-LDL和ox-LDL均可增强培养的HUVEC表达VCAM-1,尤以ox-LDL作用更明显。结论n-LDL、ox-LDL可能通过促进血管内皮细胞表达VCAM-1而在动脉粥样硬化的早期事件中起作用。 相似文献
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Summary Human arterial endothelial cells were cultured in vitro for up to 40 cumulative population doublings. Culture conditions similar
to those required for long-term propagation of human umbilical vein endothelial cells were employed. These included fibronectin-coated
culture vessels, 5 to 20% fetal bovine serum, endothelial cell growth factor, and heparin. Thoracic aorta endothelial cells
were larger than iliac artery endothelial cells. Both cell types stained positively for Factor VIII antigen by immunofluorescence.
A decrease in confluent density as a function of population doubling level was correlated with the appearance of large, senescent
cells in the cultures. Serum growth factors to which the arterial endothelial cells responded included insulin, transferrin,
epidermal growth factor, thrombin, and somatomedins. The effect of thrombin did not require the availabilty of the active
site of the protease. The effect of the somatomedins was only seen in the presence of heparin. Neither platelet-derived growth
factor nor hydrocortisone induced arteiral endothelial cell proliferation. These growth factor responses were also observed
on the part of human umbilical vein endothelial cells.
This work was supported in part by Public Health Service grants HL01030, HL01734, and AG00599. 相似文献
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Combined non-radioactive detection of peptide hormones and their mRNAs in stomach somatostatin cells
Non-radioactive in situ hybridization (ISH) and immunocytochemistry (ICC) have been used to detect somatostatin (SS) messenger RNA (mRNA) and peptide in antropyloric mucosa of the stomach in the rats. We have applied a method of non-radioactive in situ hybridization histochemistry using digoxigenin labelled oligonucleotide probes to detect somatostatin gene expression in the stomach. In prehybridization stage we used proteinase K (PK) in various concentrations (from 1 to 10 micrograms/ml) and periods (from 10 min to 1 h) but we maintained high background. However it was possible to detect the somatostatin mRNAs in the stomach mucosa making use of either background preventing solutions during the prehybridization, or of levamisole (20 microliters/mg) added into the hybridization buffer or of pepsin. Somatostatin mRNA and peptide signals were scattered all through the mucosa especially localized particularly at the base of the pyloric glands. SS peptide shown by ICC and SS mRNA shown by ISH were observed in different cells. 相似文献
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The expression and distribution of mRNA encoding preproatrial natriuretic peptide (ppANP) in rat brain has been investigated by in situ hybridization of two 35S-labeled synthetic DNA oligonucleotides, based on a cDNA clone sequence that encodes rat ppANP. The highest relative concentrations of ppANP mRNA were detected in the medial preoptic hypothalamic nucleus ("anteroventral/third ventricle region") and the medial habenula. Moderate concentrations of ppANP mRNA were observed in the CA1 pyramidal cells of the hippocampus, the endopiriform nucleus, the arcuate nucleus, the zona incerta, and cells of the pontine tegmental and peduculopontine nuclei. Several of these regions, including the habenula and the hypothalamic areas, have previously been reported to contain atrial natriuretic peptide (ANP)-like immunoreactivity, but the expression of ppANP mRNA in CA1 pyramidal cells suggests the occurrence of differential translation of ppANP mRNA into protein product in different brain regions, or the existence of different immunological forms of the peptide. The abundance of ppANP mRNA in brain was relatively low in comparison with that previously reported for many other mRNA species encoding other brain neuropeptides. These results demonstrate that ANP gene expression occurs in discrete neuronal populations of the CNS and that studies of the regulation of this expression should now be possible using quantitative in situ hybridization. 相似文献
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Furuhata S Ando K Oki M Aoki K Ohnishi S Aoyagi K Sasaki H Sakamoto H Yoshida T Ohnami S 《Molecular and cellular biochemistry》2007,298(1-2):125-138
Among the many tissue stem or progenitor cells recently being unveiled, endothelial progenitor cells (EPCs) have attracted
particular attention, not only because of their cardinal role in vascular biology and embryology but also because of their
potential use in the therapeutic development of a variety of postnatal diseases, including cardiovascular and peripheral vascular
disorders and cancer. The aim of this study is to provide some basic and comprehensive information on gene expression of EPCs
to characterize the cells in molecular terms. Here, we focus on EPCs derived from CD34-positive mononuclear cells of human
umbilical cord blood. The EPCs were purified and expanded in culture and analyzed by a high-density oligonucleotide microarray
and real-time RT-PCR analysis. We identified 169 up-regulated and 107 down-regulated genes in the EPCs compared with three
differentiated endothelial cells of human umbilical vein endothelial cells (HUVEC), human lung microvascular endothelial cells
(LMEC) and human aortic endothelial cells (AoEC). It is expected that the obtained list include key genes which are critical
for EPC function and survival and thus potential targets of EPC recognition in vivo and therapeutic modulation of vasculogenesis
in cancer as well as other diseases, in which de novo vasculogenesis plays a crucial role. For instance, the list includes
Syk and galectin-3, which encode protein tyrosine kinase and β-galactoside-binding protein, respectively, and are expressed higher in EPCs than
the three control endothelial cells. In situ hybridization showed that the genes were expressed in isolated cells in the fetal
liver at E11.5 and E14.5 of mouse development. 相似文献