血管内皮生长因子对缺血性脑血管病血管生成的作用研究

血管内皮生长因子是血管特异性生长因子,有促进内皮细胞增生、转移,增加血管通透性和促进血管生成的作用.阐述了血管内皮生长因子的生物学特性和功能,以及血管内皮生长因子与缺血性脑血管病的关系.     

An inducible endothelial cell surface glycoprotein mediates melanoma adhesion

Hematogenous metastasis requires the arrest and extravasation of blood-borne tumor cells, possibly involving direct adhesive interactions with vascular endothelium. Cytokine activation of cultured human endothelium increases adhesion of melanoma and carcinoma cell lines. An inducible 110-kD endothelial cell surface glycoprotein, designated INCAM-110, appears to mediate adhesion of melanoma cells. In addition, an inducible endothelial receptor for neutrophils, ELAM-1, supports the adhesion of a human colon carcinoma cell line. Thus, activation of vascular endothelium in vivo that results in increased expression of INCAM-110 and ELAM-1 may promote tumor cell adhesion and affect the incidence and distribution of metastases.     

G protein-coupled receptor-dependent development of human frontal cortex

The mammalian cerebral cortex is characterized by complex patterns of anatomical and functional areas that differ markedly between species, but the molecular basis for this functional subdivision is largely unknown. Here, we show that mutations in GPR56, which encodes an orphan G protein-coupled receptor (GPCR) with a large extracellular domain, cause a human brain cortical malformation called bilateral frontoparietal polymicrogyria (BFPP). BFPP is characterized by disorganized cortical lamination that is most severe in frontal cortex. Our data suggest that GPCR signaling plays an essential role in regional development of human cerebral cortex.     

Angiogenesis-independent endothelial protection of liver: role of VEGFR-1

The vascular endothelium was once thought to function primarily in nutrient and oxygen delivery, but recent evidence suggests that it may play a broader role in tissue homeostasis. To explore the role of sinusoidal endothelial cells (LSECs) in the adult liver, we studied the effects of vascular endothelial growth factor (VEGF) receptor activation on mouse hepatocyte growth. Delivery of VEGF-A increased liver mass in mice but did not stimulate growth of hepatocytes in vitro, unless LSECs were also present in the culture. Hepatocyte growth factor (HGF) was identified as one of the LSEC-derived paracrine mediators promoting hepatocyte growth. Selective activation of VEGF receptor-1 (VEGFR-1) stimulated hepatocyte but not endothelial proliferation in vivo and reduced liver damage in mice exposed to a hepatotoxin. Thus, VEGFR-1 agonists may have therapeutic potential for preservation of organ function in certain liver disorders.     

Role of the sphingosine-1-phosphate receptor EDG-1 in PDGF-induced cell motility

EDG-1 is a heterotrimeric guanine nucleotide binding protein-coupled receptor (GPCR) for sphingosine-1-phosphate (SPP). Cell migration toward platelet-derived growth factor (PDGF), which stimulates sphingosine kinase and increases intracellular SPP, was dependent on expression of EDG-1. Deletion of edg-1 or inhibition of sphingosine kinase suppressed chemotaxis toward PDGF and also activation of the small guanosine triphosphatase Rac, which is essential for protrusion of lamellipodia and forward movement. Moreover, PDGF activated EDG-1, as measured by translocation of beta-arrestin and phosphorylation of EDG-1. Our results reveal a role for receptor cross-communication in which activation of a GPCR by a receptor tyrosine kinase is critical for cell motility.     

中医药调控内皮祖细胞的现状与思路

内皮祖细胞(endothelial progenitor cell,EPC)是修复损伤血管内皮的细胞库,EPC功能异常和数量减少是内皮功能障碍的主要原因,而内皮功能障碍是多种内皮损伤性疾病的始动因素。利用细胞疗法输注体外扩增的EPC来修复受损血管内皮已成为当前防治心脑血管疾病的有效手段。但单独输注EPC修复受损血管内皮的疗效不甚明显,其与血管内皮损伤后局部微环境的改变以及多种细胞因子的干预影响EPC的动员、归巢和分化有较大关系。因此探讨药物干预促进EPC对受损血管内皮的修复具有一定的科学意义,而中医药是伟大的宝库,从中选择有效的药物和方法促进EPC修复受损血管内皮具有广阔的应用前景。     

Function of PI3Kgamma in thymocyte development, T cell activation, and neutrophil migration

Phosphoinositide 3-kinases (PI3Ks) regulate fundamental cellular responses such as proliferation, apoptosis, cell motility, and adhesion. Viable gene-targeted mice lacking the p110 catalytic subunit of PI3Kgamma were generated. We show that PI3Kgamma controls thymocyte survival and activation of mature T cells but has no role in the development or function of B cells. PI3Kgamma-deficient neutrophils exhibited severe defects in migration and respiratory burst in response to heterotrimeric GTP-binding protein (G protein)-coupled receptor (GPCR) agonists and chemotactic agents. PI3Kgamma links GPCR stimulation to the formation of phosphatidylinositol 3,4,5-triphosphate and the activation of protein kinase B, ribosomal protein S6 kinase, and extracellular signal-regulated kinases 1 and 2. Thus, PI3Kgamma regulates thymocyte development, T cell activation, neutrophil migration, and the oxidative burst.     

High-level recombinant gene expression in rabbit endothelial cells transduced by retroviral vectors

By virtue of its immediate contact with the circulating blood, the endothelium provides an attractive target for retroviral vector transduction for the purpose of gene therapy. To see whether efficient gene transfer and expression was feasible, rabbit aortic endothelial cells were infected with three Moloney murine leukemia virus-derived retroviral vectors. Two of these vectors carry genes encoding products that are not secreted: N2, containing only the selectable marker gene neoR, and SAX, containing both neoR gene and an SV40-promoted adenosine deaminase (ADA) gene. The third vector, G2N, contains a secretory rat growth hormone (rGH) gene and an SV40-promoted neoR gene. Infection with all three vectors resulted in expression of the respective genes. A high level of human ADA expression was observed in infected endothelial cell populations both before and after selection in G418. G2N-infected rabbit aortic endothelial cells that were grown on a synthetic vascular graft continued to secrete rGH into the culture medium. These studies suggest that endothelial cells may serve as vehicles for the introduction in vivo of functioning recombinant genes.     

Structurally homologous ligand binding of integrin Mac-1 and viral glycoprotein C receptors

Three spatially distant surface loops were found to mediate the interaction of the coagulation protein factor X with the leukocyte integrin Mac-1. This interacting region, which by computational modeling defines a three-dimensional macromotif in the catalytic domain, was also recognized by glycoprotein C (gC), a factor X receptor expressed on herpes simplex virus (HSV)-infected endothelial cells. Peptidyl mimicry of each loop inhibited factor X binding to Mac-1 and gC, blocked monocyte generation of thrombin, and prevented monocyte adhesion to HSV-infected endothelium. These data link the ligand recognition of Mac-1 to established mechanisms of receptor-mediated vascular injury.     

Elementary Ca2+ signals through endothelial TRPV4 channels regulate vascular function

Major features of the transcellular signaling mechanism responsible for endothelium-dependent regulation of vascular smooth muscle tone are unresolved. We identified local calcium (Ca(2+)) signals ("sparklets") in the vascular endothelium of resistance arteries that represent Ca(2+) influx through single TRPV4 cation channels. Gating of individual TRPV4 channels within a four-channel cluster was cooperative, with activation of as few as three channels per cell causing maximal dilation through activation of endothelial cell intermediate (IK)- and small (SK)-conductance, Ca(2+)-sensitive potassium (K(+)) channels. Endothelial-dependent muscarinic receptor signaling also acted largely through TRPV4 sparklet-mediated stimulation of IK and SK channels to promote vasodilation. These results support the concept that Ca(2+) influx through single TRPV4 channels is leveraged by the amplifier effect of cooperative channel gating and the high Ca(2+) sensitivity of IK and SK channels to cause vasodilation.     

Role of Raf in vascular protection from distinct apoptotic stimuli

Raf kinases have been linked to endothelial cell survival. Here, we show that basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF) differentially activate Raf, resulting in protection from distinct pathways of apoptosis in human endothelial cells and chick embryo vasculature. bFGF activated Raf-1 via p21-activated protein kinase-1 (PAK-1) phosphorylation of serines 338 and 339, resulting in Raf-1 mitochondrial translocation and endothelial cell protection from the intrinsic pathway of apoptosis, independent of the mitogen-activated protein kinase kinase-1 (MEK1). In contrast, VEGF activated Raf-1 via Src kinase, leading to phosphorylation of tyrosines 340 and 341 and MEK1-dependent protection from extrinsic-mediated apoptosis. These findings implicate Raf-1 as a pivotal regulator of endothelial cell survival during angiogenesis.     

枸杞多糖组分-Ⅳ对有机磷农药所致血管内皮功能损伤的保护作用

[目的]探讨枸杞多糖组分-Ⅳ（LBP-Ⅳ）对对氧磷（PARA）所致血管内皮功能损伤的保护作用及其机制。[方法]以大鼠离体胸主动脉血管环（EVAPVR）和培养的人脐静脉内皮细胞（HUVEC）为试验对象,以PARA为损伤药物,以LBP-Ⅳ为保护药,检测血管内皮依赖性舒张反应（EDRR）、内皮细胞单层通透性（ECMP）及细胞培养液生化指标。[结果]LBP-Ⅳ剂量依赖性（0.1、1、10mg/ml）地显著减轻了PARA（3.63μmol/l）对血管EDRR的抑制作用,降低了ECMP的增加,保护了超氧化物歧化酶（SOD）活性,阻滞了丙二醛（MDA）浓度的升高以及一氧化氮（NO）浓度的降低（P〈0.05）。[结论]LBP-Ⅳ对PARA所致的血管内皮功能损伤有明显的保护作用,其机制可能与LBP-Ⅳ的抗氧化作用有关。     

黄芪多糖组分-A1对甲基异柳磷所致血管内皮功能损伤的保护作用

[目的]研究黄芪多糖组分-A1(APS-A1)对甲基异柳磷所致血管内皮功能损伤的保护作用。[方法]以大鼠离体胸主动脉血管环(EVAPVR)和培养的人脐静脉内皮细胞(HUVEC)为试验对象,以甲基异柳磷为损伤药物,以APS-A1为保护药,检测血管内皮依赖性舒张反应(EDRR)、内皮细胞单层通透性(ECMP)及细胞培养液生化指标。[结果]APS-A1能明显减轻甲基异柳磷(7μmol/L)对血管EDRR的抑制作用,且呈剂量依赖性;APS-A1也能降低ECMP的增加,保护超氧化物歧化酶(SOD)的活性,并阻滞丙二醛(MDA)浓度的升高以及一氧化氮(NO)浓度的降低。[结论]APS-A1对甲基异柳磷所致的血管内皮功能损伤有明显的保护作用。     

Comment on "Obestatin, a peptide encoded by the ghrelin gene, opposes ghrelin's effects on food intake"

Zhang et al. (Research Articles, 11 November 2005, p. 996) reported that obestatin, a peptide derived from the ghrelin precursor, activated the orphan G protein-coupled receptor GPR39. However, we found that I125-obestatin does not bind GPR39 and observed no effects of obestatin on GPR39-transfected cells in various functional assays (cyclic adenosine monophosphate production, calcium mobilization, and GPR39 internalization). Our results indicate that obestatin is not the cognate ligand for GPR39.     

Vascular endothelial growth factor is a secreted angiogenic mitogen

Vascular endothelial growth factor (VEGF) was purified from media conditioned by bovine pituitary folliculostellate cells (FC). VEGF is a heparin-binding growth factor specific for vascular endothelial cells that is able to induce angiogenesis in vivo. Complementary DNA clones for bovine and human VEGF were isolated from cDNA libraries prepared from FC and HL60 leukemia cells, respectively. These cDNAs encode hydrophilic proteins with sequences related to those of the A and B chains of platelet-derived growth factor. DNA sequencing suggests the existence of several molecular species of VEGF. VEGFs are secreted proteins, in contrast to other endothelial cell mitogens such as acidic or basic fibroblast growth factors and platelet-derived endothelial cell growth factor. Human 293 cells transfected with an expression vector containing a bovine or human VEGF cDNA insert secrete an endothelial cell mitogen that behaves like native VEGF.     

Leakage-resistant blood vessels in mice transgenically overexpressing angiopoietin-1

Angiopoietin-1 (Ang1) and vascular endothelial growth factor (VEGF) are endothelial cell-specific growth factors. Direct comparison of transgenic mice overexpressing these factors in the skin revealed that the VEGF-induced blood vessels were leaky, whereas those induced by Ang1 were nonleaky. Moreover, vessels in Ang1-overexpressing mice were resistant to leaks caused by inflammatory agents. Coexpression of Ang1 and VEGF had an additive effect on angiogenesis but resulted in leakage-resistant vessels typical of Ang1. Ang1 therefore may be useful for reducing microvascular leakage in diseases in which the leakage results from chronic inflammation or elevated VEGF and, in combination with VEGF, for promoting growth of nonleaky vessels.     

Enhanced morphine analgesia in mice lacking beta-arrestin 2

The ability of morphine to alleviate pain is mediated through a heterotrimeric guanine nucleotide binding protein (G protein)-coupled heptahelical receptor (GPCR), the mu opioid receptor (muOR). The efficiency of GPCR signaling is tightly regulated and ultimately limited by the coordinated phosphorylation of the receptors by specific GPCR kinases and the subsequent interaction of the phosphorylated receptors with beta-arrestin 1 and beta-arrestin 2. Functional deletion of the beta-arrestin 2 gene in mice resulted in remarkable potentiation and prolongation of the analgesic effect of morphine, suggesting that muOR desensitization was impaired. These results provide evidence in vivo for the physiological importance of beta-arrestin 2 in regulating the function of a specific GPCR, the muOR. Moreover, they suggest that inhibition of beta-arrestin 2 function might lead to enhanced analgesic effectiveness of morphine and provide potential new avenues for the study and treatment of pain, narcotic tolerance, and dependence.     

Angiogenic factors

Within the past 2 years, several angiogenic factors have been fully purified, their amino acid sequences determined, and their genes cloned. These polypeptides include acidic and basic fibroblast growth factor, angiogenin, and transforming growth factors alpha and beta. Other less well characterized angiogenesis factors have also been isolated, some of which are lipids. This article traces the discovery of the angiogenic factors and describes their possible significance in understanding growth regulation of the vascular system. When evaluated according to their putative targets, they appear to fall into two groups: those that act directly on vascular endothelial cells to stimulate locomotion or mitosis, and those that act indirectly by mobilizing host cells (for example, macrophages) to release endothelial growth factors. In addition to their presence in tumors undergoing neovascularization, the same angiogenic peptides are found in many normal tissues where neovascularization is not occurring. This suggests that physiological expression of angiogenic factors is tightly regulated. In addition to the persistent angiogenesis induced by tumors, it now appears that a variety of nonneoplastic diseases, previously thought to be unrelated, can be considered as "angiogenic diseases" because they are dominated by the pathologic growth of capillary blood vessels.     

辛伐他丁对2型糖尿病大鼠血管内皮VCAM-1和MCP-1mRNA和蛋白表达的影响

目的了解辛伐他丁对2型糖尿病大鼠血管内皮细胞黏附分子-1(VCAM-1)和单核细胞趋化因子-1(MCP-1)的影响。方法30只SD大鼠随机分为正常组(NC)、糖尿病组(DMC)、辛伐他丁治疗组(DMT),每组各10只大鼠。喂养12周后抽血测定血脂全套,取主动脉标本以免疫组化和半定量RT-PCR法分别检测VCAM-1和MCP-1的mRNA和蛋白表达。结果DMT组大鼠血管内皮VCAM-1和MCP-1的mRNA和蛋白表达明显降低,与DMC组比较差异有统计学意义(P<0.05)。结论辛伐他丁可减少糖尿病大鼠血管内皮VCAM-1和MCP-1的表达,减轻血管内皮局部的炎症反应。