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.     

Vascular abnormalities and deregulation of VEGF in Lkb1-deficient mice

The LKB1 tumor suppressor gene, mutated in Peutz-Jeghers syndrome, encodes a serine/threonine kinase of unknown function. Here we show that mice with a targeted disruption of Lkb1 die at midgestation, with the embryos showing neural tube defects, mesenchymal cell death, and vascular abnormalities. Extraembryonic development was also severely affected; the mutant placentas exhibited defective labyrinth layer development and the fetal vessels failed to invade the placenta. These phenotypes were associated with tissue-specific deregulation of vascular endothelial growth factor (VEGF) expression, including a marked increase in the amount of VEGF messenger RNA. Moreover, VEGF production in cultured Lkb1(-/-) fibroblasts was elevated in both normoxic and hypoxic conditions. These findings place Lkb1 in the VEGF signaling pathway and suggest that the vascular defects accompanying Lkb1 loss are mediated at least in part by VEGF.     

齐墩果酸抗大鼠移植性肝癌作用的研究

[目的]研究齐墩果酸抗肿瘤作用以及对肿瘤血管内皮生长因子（VEGF）和微血管密度（MVD）的影响,探讨齐墩果酸抗肝癌作用机制。[方法]32只大鼠移植性肝癌模型随机分为齐墩果酸高剂量组（100mS／kg·d）、齐墩果酸低剂量组（50mg／kg·d）、氟尿嘧啶（5-Fu）组（5mg／kg·d）、生理盐水组,每组8只,前3组于造模成功后第8天腹腔注射给药,生理盐水组灌胃给药,连续用药14d后停药24h,取血处死大鼠,取出瘤块称重,常规瘤体组织形态学观察,分离血清检测肝功能指标谷草转氨酶（AST）、谷丙转氨酶（ALT）,用免疫组化法检测瘤块中VEGF和MVD的表达情况。[结果1齐墩果酸给药组瘤重明显低于生理盐水对照组,与生理盐水对照组比较差异显著（P〈0．01）;齐墩果酸给药组及5一FIJ组血管条数目密集程度较生理盐水组明显减少;齐墩果酸对血清中AsT、AIJT活性升高有明显的拮抗作用,并且AsT的含量与生理盐水对照组比较有显著性差异（P〈0．01）;造模各组VEGF及MVD的表达水平均明显高于空白组（P〈0．01）,两者的表达均以生理盐水组最高。l结论]齐墩果酸可显著抑制大鼠体内癌细胞的生长,对移植性肝癌所造成的肝功能损伤具有明显的保护作用,其作用机制可能与下调VEGF和MVD的表达水平有关。     

VEGF和metastin的表达与乳腺癌临床病理的关系

目的分析血管内皮生长因子(VEGF)和KISS-1基因编码产物metastin与乳腺癌生物学行为的关系。方法采用免疫组化SP法观察70例乳腺癌组织和癌周正常组织中VEGF和metastin的表达情况。结果VEGF在乳腺癌组织中的阳性表达率为88.6%(62/70),在癌旁正常组织中的阳性表达率为42.9%(30/70),两者比较差异有统计学意义(P<0.01);metastin在乳腺癌组织中的阳性表达率为30.0%(21/70),在癌旁正常组织中的阳性表达率为72.9%(51/70),两者比较差异有统计学意义(P<0.01)。VEGF表达水平与乳腺癌肿瘤大小、TNM分期、淋巴结转移有关(P<0.05),metastin表达水平与乳腺癌TNM分期、淋巴结转移有关(P<0.05)。结论VEGF和metastin在乳腺癌发生、发展过程中发挥了一定的作用,VEGF与metastin表达水平的同时检测,对分析乳腺癌恶性程度和预后有重要意义。     

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.     

血管内皮生长因子研究进展

血管内皮生长因子(Vascular Endothelial Growth Factor, VEGF)是一种由各种正常细胞或肿瘤细胞合成和分泌的糖蛋白。人类医学研究证明,VEGF有两种受体——KDR/FLK-1和FLT-1。VEGF与受体结合,具有促进血管内皮细胞的增生和提高微血管对大分子物质的通透性作用。人的VEGF 基因由8个外显子构成,主要有VEGF121、VEGF145、VEGF165、VEGF189和VEGF206等5个同型异构体。等电点的差异和肝素亲合力的差异可能是造成VEGF 生物活性差异的主要原因。     

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.     

血管内皮生长因子在上皮性卵巢肿瘤中的表达及其意义

目的了解血管内皮生长因子(vascular endothelial growth factor,VEGF)在上皮性卵巢肿瘤中的表达及其意义。方法应用ELISA法检测上皮性卵巢癌患者(恶性组,n=30)、卵巢良性肿瘤患者(良性组,n=30)和正常妇女(对照组,n=20)外周血中VEGF水平。结果恶性组血清VEGF水平明显高于良性组和对照组(P<0.01),良性组和对照组相比较差异无统计学意义(P>0.05)。恶性组术前血清VEGF水平明显高于术后(P<0.01),良性组术前、术后的VEGF水平差异无统计学意义(P>0.05)。恶性组Ⅲ、Ⅳ期患者的VEGF水平高于Ⅰ、Ⅱ期,低分化G3期的VEGF水平高于高中分化G1、2期,差异有统计学意义(P<0.01)。结论检测血清VEGF水平对上皮性卵巢肿瘤的诊断及恶性上皮性肿瘤的分期、分级和预后有重要意义。     

IL-8基因沉默后猪血管内皮细胞相关细胞因子表达的变化

为了探究内皮源IL-8对猪血管内皮细胞其他相关细胞因子表达的影响,解析IL-8在内皮细胞中作用,利用siRNA干扰技术对猪血管内皮细胞（VEC）中IL-8基因进行沉默,于干扰后不同时间收集细胞及上清,应用荧光定量PCR技术和ELISA检测VEC中各细胞因子的变化.结果显示,在mRNA水平上,促炎因子IL-6和促内皮生长因子（VEGF）在12h上调显著,且在4 h VEGF显著上调;ICAM-1和VCAM-1在4h显著上调,其中ICAM-1在48 h极显著下调,72 h又显著上调;巨噬细胞集落刺激因子（M-CSF）几个时间段均极显著高于对照组.蛋白水平上,VEGF除48 h外均显著高于对照组,M-CSF在4h后均显著低于对照组.以上数据表明,内皮源IL-8变化可能影响VEC增殖、迁移以及对造血干细胞分化.     

羊膜移植抑制碱烧伤后角膜血管内皮生长因子表达的实验研究

目的探讨羊膜移植对角膜内血管内皮生长因子(vascular endothelial growth factor,VEGF)及角膜新生血管形成的影响,方法12只兔(24眼)被建立角膜碱烧伤动物模型,行羊膜移植的左眼为实验组,右眼为对照组,分别采用免疫组化染色检查VEGF蛋白的表达,宏观测量新生血管长度及生长速度,显微镜下微血管计数方法研究角膜新生血管形成及抑制情况,结果与对照组比较,实验组兔眼角膜的VEGF蛋白表达下降,新生血管生长速度变慢,微血管数量减少(P＜0．01),VEGF主要表达在角膜受损区的炎性细胞胞浆内,其出现时间及位置与角膜新生血管一致,结论VEGF是一种重要的角膜内新生血管形成因子,其变化与角膜新生血管平行;羊膜移植可抑制碱烧伤角膜内VEGF的表达及角膜新生血管的形成.     

Syk和VEGF在胃癌组织中的表达及其临床意义

目的观察脾酪氨酸激酶（Syk）和血管内皮生长因子（VEGF）在胃癌组织中的表达情况,分析它们与胃癌病理临床特征之间的关系。方法应用免疫组化法检测100例胃癌患者手术切除的癌组织和癌周正常组织中Syk和VEGF表达水平。结果Syk在正常胃组织中表达率为100．0％（100／100）,在胃癌组织中的表达率为40．0％（40／100）;VEGF在正常胃组织中表达率为0．0％（0／100）,在胃癌组织中的表达率为50．0％（50／100）,两者比较差异均有统计学意义（P〈0．01）。Syk蛋白表达水平与胃癌浆膜浸润、肿瘤大小、TNM分期、淋巴结转移有关（P〈0．05）,VEGF蛋白表达水平与胃癌浆膜浸润、TNM分期、淋巴结转移有关（P〈0．05）。结论同时检测胃癌组织中Syk与VEGF表达水平,对判断胃癌浸润转移潜能及评估患者预后有一定的参考价值。     

Regulation of hepatic stellate cell differentiation by the neurotrophin receptor p75NTR

Differentiation of hepatic stellate cells (HSCs) to extracellular matrix- and growth factor-producing cells supports liver regeneration through promotion of hepatocyte proliferation. We show that the neurotrophin receptor p75NTR, a tumor necrosis factor receptor superfamily member expressed in HSCs after fibrotic and cirrhotic liver injury in humans, is a regulator of liver repair. In mice, depletion of p75NTR exacerbated liver pathology and inhibited hepatocyte proliferation in vivo. p75NTR-/- HSCs failed to differentiate to myofibroblasts and did not support hepatocyte proliferation. Moreover, inhibition of p75NTR signaling to the small guanosine triphosphatase Rho resulted in impaired HSC differentiation. Our results identify signaling from p75NTR to Rho as a mechanism for the regulation of HSC differentiation to regeneration-promoting cells that support hepatocyte proliferation in the diseased liver.     

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.     

梅花鹿VEGF基因的克隆及其在鹿茸顶端组织的表达

为了得到VEGF成熟肽基因,利用Trizol试剂法提取鹿茸顶端组织总RNA,反转录形成c DNA。根据Gen Bank已发表的相关基因序列设计1对特异性引物,并克隆VEGF成熟肽基因,再利用免疫组化法检测VEGF基因在鹿茸顶端不同组织的表达水平。结果表明:成功获得梅花鹿VEGF成熟肽基因,该基因长495 bp,共编码164个氨基酸;经Blast同源性分析,结果显示与牛、羊、猪的核苷酸序列的同源性分别为98.79%、97.98%、96.36%;经DNAMAN软件比对,其氨基酸序列的同源性分别为98.78%、97.56%、96.95%。免疫组化法结果显示,VEGF在茸皮层、间充质层和软骨层中均有表达,其中在软骨层中表达水平较高。     

斑马鱼不同发育时期血管内皮生长因子受体2基因表达的实时定量PCR分析

[目的]探讨血管内皮生长因子受体2（VEGFR2-）基因在斑马鱼不同发育时期的表达。[方法]分别从12、24、48、72和96 h的斑马鱼胚胎和仔鱼中提取总RNA,用实时定量RT-PCR方法检测VEGFR2-基因表达,采用2^-△△Ct法进行数据分析。[结果]VEGFR2-基因表达量在12-72 h呈上升趋势,在96 h有所下降。12 h表达量最低,72 h表达量最高,与其他发育期均有显著差异。[结论]斑马鱼血管发育至72 h达成熟阶段。血管发育成熟前,VEGFR2-基因表达水平逐步增长;血管发育成熟后,VEGFR2-基因表达水平下降。     

LRP: role in vascular wall integrity and protection from atherosclerosis

Vascular smooth muscle cell (SMC) proliferation and migration are important events in the development of atherosclerosis. The low-density lipoprotein receptor-related protein (LRP1) mediates suppression of SMC migration induced by platelet-derived growth factor (PDGF). Here we show that LRP1 forms a complex with the PDGF receptor (PDGFR). Inactivation of LRP1 in vascular SMCs of mice causes PDGFR overexpression and abnormal activation of PDGFR signaling, resulting in disruption of the elastic layer, SMC proliferation, aneurysm formation, and marked susceptibility to cholesterol-induced atherosclerosis. The development of these abnormalities was reduced by treatment with Gleevec, an inhibitor of PDGF signaling. Thus, LRP1 has a pivotal role in protecting vascular wall integrity and preventing atherosclerosis by controlling PDGFR activation.     

Liver organogenesis promoted by endothelial cells prior to vascular function

The embryonic role of endothelial cells and nascent vessels in promoting organogenesis, prior to vascular function, is unclear. We find that early endothelial cells in mouse embryos surround newly specified hepatic endoderm and delimit the mesenchymal domain into which the liver bud grows. In flk-1 mutant embryos, which lack endothelial cells, hepatic specification occurs, but liver morphogenesis fails prior to mesenchyme invasion. We developed an embryo tissue explant system that permits liver bud vasculogenesis and show that in the absence of endothelial cells, or when the latter are inhibited, there is a selective defect in hepatic outgrowth. We conclude that vasculogenic endothelial cells and nascent vessels are critical for the earliest stages of organogenesis, prior to blood vessel function.     

Role of prostacyclin in the cardiovascular response to thromboxane A2

Thromboxane (Tx) A2 is a vasoconstrictor and platelet agonist. Aspirin affords cardioprotection through inhibition of TxA2 formation by platelet cyclooxygenase (COX-1). Prostacyclin (PGI2) is a vasodilator that inhibits platelet function. Here we show that injury-induced vascular proliferation and platelet activation are enhanced in mice that are genetically deficient in the PGI2 receptor (IP) but are depressed in mice genetically deficient in the TxA2 receptor (TP) or treated with a TP antagonist. The augmented response to vascular injury was abolished in mice deficient in both receptors. Thus, PGI2 modulates platelet-vascular interactions in vivo and specifically limits the response to TxA2. This interplay may help explain the adverse cardiovascular effects associated with selective COX-2 inhibitors, which, unlike aspirin and nonsteroidal anti-inflammatory drugs (NSAIDs), inhibit PGI2 but not TxA2.