Ang-2在大肠癌血管生成和肿瘤生长中的作用

目的探讨促血管生成素Ⅱ(Angiopoietin-2，Ang-2)在肠腺瘤和大肠癌组织生长和血管生成中的作用。方法采用免疫组化实验方法检测62例大肠癌和30例肠腺瘤手术标本中Ang-2、CD-34和ki67蛋白表达量，同时以16例正常大肠粘膜组织作对照。结果Ang-2、ki67蛋白和微血管密度(MVD)在大肠癌和肠腺瘤组织中均有高表达，正常大肠粘膜组织表达量少；肠癌组织表达水平明显高于肠腺瘤样组织，且两者均高于正常对照组；肠癌Ang-2与MVD和ki67在相应组织中的表达成正相关。结论肠癌和肠腺瘤组织中过多表达Ang-2与肿瘤血管和肿瘤生长密切相关，Ang-2在大肠癌血管生成及其肿瘤发生、发展过程中起着重要作用。     

Mizolastine（MIZ）对趾叶炎小鼠趾部组织VEGF表达的影响

用组织化学和免疫组化的染色方法对趾叶炎发生过程中正常组、造模组和低、中、高剂量治疗组小鼠趾部组织血管内皮生长因子（VEGF）表达水平、血管数量和肥大细胞及其脱颗粒的动态变化进行了研究.结果表明,造模组小鼠在免疫后14d,VEGF表达均达到一个峰值,而后表达开始减弱,21d后表达开始增强,均极显著高于同一时相的正常组（P〈0.01）.低、中、高剂量治疗组的VEGF表达均极显著低于同一时相造模组,其中,低剂量治疗组同一时相均极显著高于正常组（P〈0.01）,中剂量治疗组除免疫后前3个时相显著外（P〈0.05）,免疫后35d与正常组不显著（P〉0.05）,高剂量治疗组除免疫后14d显著外（P〈0.05）,其余各时相均与正常组不显著（P〉0.05）,治疗各组小鼠的VEGF表达水平随药物浓度的增大和免疫时间的延长而逐渐减弱;真皮内出现了许多增生的小动脉,管壁增厚,管腔内有血栓形成,且血管形成与肥大细胞及其脱颗粒的变化均与VEGF的表达趋势相吻合.Mizo-lastine（MIZ）可以显著抑制肥大细胞脱颗粒、VEGF的表达和血管形成.     

微血管密度与血管内皮生长因子受体-3在骨髓增生异常综合征患者中的表达及意义

目的检测骨髓增生异常综合征患者骨髓中微血管密度和血管内皮生长因子受体-3表达水平并探讨其临床意义。方法采用免疫组织化学染色检测54例骨髓增生异常综合症患者、20例非恶性血液病患者对照组骨髓组织中微血管密度和血管内皮生长因子受体-3表达水平,采用蛋白质印迹法检测上述研究对象骨髓单个核细胞中血管内皮生长因子受体-3蛋白表达水平,分析其与临床特征的相关性。结果骨髓增生异常综合症患者骨髓组织中微血管密度和血管内皮生长因子受体-3表达水平明显高于非恶性血液病对照组,差异具有统计学意义(P0.05),而骨髓增生异常综合症低危组、中危组及高危组中微血管密度与血管内皮生长因子受体-3的表达水平无显著差异(P0.05),经相关性分析微血管密度和血管内皮生长因子受体-3在骨髓增生异常综合症患者骨髓中表达呈正相关关系。结论骨髓增生异常综合症患者骨髓存在明显血管新生及血管内皮生长因子受体-3高表达,其表达水平可能参与骨髓增生异常综合症发病、发展等过程,并可能影响骨髓增生异常综合症预后,通过检测微血管密度和血管内皮生长因子受体-3的表达可为临床抗脉管新生方法治疗骨髓增生异常综合症提供实验依据。     

血管内皮生长因子在大鼠乳腺炎中的作用

应用组织化学方法及免疫组化技术对正常组和急性攻毒组大鼠乳腺组织血管内皮生长因子（Vascular Endothelial Growth Factor,VEGF）的表达水平、肥大细胞的活力和炎性细胞的动态变化进行了研究.结果表明,急性攻毒后6 h乳腺组织VEGF在腺泡间、小叶间质、血管周围及部分肥大细胞内的表达出现一个峰值,而后表达开始减弱,攻毒48 h后表达又迅速增强;攻毒组大鼠的肥大细胞及其脱颗粒和炎性细胞随着攻毒时间的延长急剧增多,均极显著高于正常组（P〈0.01）,且其变化趋势均与VEGF表达趋势相吻合;同时,小叶间质内的小血管内皮细胞开始出现裂隙、脱落损伤.由此表明,VEGF和肥大细胞参与大鼠乳腺炎的发病过程.     

糖尿病性勃起功能障碍多因素发病机制研究

从神经、血管、代谢、离子通道等多个角度出发,研究糖尿病性勃起功能障碍(DMED)的发病机制,为开展DMED的综合治疗临床研究提供前期研究基础.成年雄性SD大鼠50只,随机取35只大鼠用于制作糖尿病模型,其余大鼠作为正常对照.饲养8周后,用阿朴吗啡法筛选DMED大鼠,用电刺激勃起神经测定海绵体内压(ICP)方法评价勃起功能.取正常组和DMED组大鼠的阴茎海绵体组织,分成若干份,用免疫组化法测定海绵体组织一氧化氮合酶(NOS)的3种亚型(nNOS,eNOS,iNOS)的变化;用Western Blot法测定神经生长因子(NGF)在海绵体组织的表达差异;用放射免疫法测定海绵体组织血管紧张素II(AngII)含量差异;用Western Blot法测定血管内皮生长因子(VEGF)和缝隙连接蛋白Connexin-43含量差异.与正常对照组相比,DMED大鼠ICP测定值明显降低;海绵体组织中nNOS、eNOS表达明显降低,而iNOS表达明显增加;NGF蛋白表达明显增加;血管紧张素II水平显著升高;VEGF蛋白表达明显减少;Connexin-43蛋白表达减少.以上试验结果说明DMED发病机制复杂,与神经、血管、代谢、离子通道等均有关.提示在今后临床治疗研究和实践中,应该从各个角度出发,针对多因素发病机制,采用综合治疗的方法,以提高疗效.     

PTEN基因在哈萨克族食管癌组织中的表达及意义

通过观察PTEN在哈萨克族食管癌癌组织及相应正常组织中的表达,探讨PTEN基因在哈萨克族食管癌发生中的作用。采用RT-PCR方法检测PTEN基因mRNA在36例哈萨克族食管癌及相应正常组织标本中的表达,选取mRNA水平差异组织标本,采用Western-bolt方法检测其蛋白水平的表达情况。结果显示,PTEN基因mRNA在36例癌组织中有30例表达,阳性率为83.33%。其中癌组织表达量高于相应正常组织(TN)21例,占70%;癌组织表达量等于相应正常组织(T=N)4例,占13.33%;癌组织表达量低于相应正常组织(TN)5例,占16.67%。PTEN基因mRNA在正常组织中表达仅有9例。癌组织与相应正常组织相比,PTEN基因的表达量明显增高。PTEN蛋白的表达在癌组织与相应正常组织中无明显差异。结果表明,PTEN基因的异常表达主要在转录水平,可能不是哈萨克族食管癌发生发展中的主导基因,其对哈萨克族食管癌发生发展过程中的具体作用有待进一步研究。     

P物质在临床型乳腺炎奶牛肝组织中的表达及意义

应用组织化学和免疫组织化学方法对正常与临床型乳腺炎奶牛肝组织中P物质的表达水平、炎性细胞以及肥大细胞的数量、活力、组化性质的变化进行了研究．结果表明，P物质在临床型乳腺炎奶牛肝组织的肝血窦、肝小叶间质、中央静脉、小叶闻动脉和静脉周围并常丰富，呈单根或多交错成网走行，表达增强．临床型乳腺炎奶牛肝组织的肥大细胞和炎性细胞也急剧增多，均极显著高于正常奶牛（P〈0．01），并与P物质的表达水平相吻合．同时，肝细胞索结构出现紊乱，肝小血管的内皮细胞出现脱落．由此表明，P物质参与临床型乳腺炎病理过程中所致肝组织损伤的病理进程．     

MSP58基因在肝癌中的表达分析

探讨MSP58基因在人类肝癌肿瘤组织及肝癌细胞系中的表达情况,应用半定量RT-PCR和Western blot方法分别检测MSP58在肝癌细胞系及肝癌肿瘤组织中的mRNA水平及蛋白水平的表达.两种肝癌细胞系HHCC、HepG2,22例肝癌癌区组织,13例正常肝组织作为对照.结果在两种肝癌细胞系中MSP58 mRNA水平和蛋白水平都有明显表达;在22例肝癌肿瘤组织中,不同级别的人肝癌肿瘤标本中mRNA及蛋白均有MSP58表达,并且MSP58的表达量随着人肝癌肿瘤恶性病理级别的增高而增高;13例正常肝组织无表达.MSP58在肝癌肿瘤组织和肝癌细胞系中均有表达,说明其可能对肝癌肿瘤的发生或发展有重要作用.     

CTGF mRNA在糖尿病大鼠心肌的表达及粉防己碱保护作用的研究

目的研究粉防己碱对糖尿病(DM)大鼠心肌的保护作用及相关机制.方法36只Wistar大鼠用链脲佐菌素(STZ)诱发糖尿病模型后,随机分为粉防己碱治疗组、苯那普利治疗组及糖尿病未治疗组,以10只正常大鼠作为对照.12周末检测血糖、血脂、心肌组织血管紧张素Ⅱ,应用RT-PCR检测心肌CTGR mRNA的表达水平.结果DM未治疗组心肌组织血管紧张素Ⅱ含量、CTGR mRNA的表达明显高于对照组和治疗组(P<0.01).结论持续高糖能导致心肌血管紧张素Ⅱ含量增高、CTGF mRNA表达增强,而粉防己碱能在一定程度上降低心肌血管紧张素Ⅱ含量,抑制CTGF mRNA表达,从而可能减轻糖尿病性心肌病的病理变化.     

Pathophysiological consequences of VEGF-induced vascular permeability

Although vascular endothelial growth factor (VEGF) induces angiogenesis, it also disrupts vascular barrier function in diseased tissues. Accordingly, VEGF expression in cancer and ischaemic disease has unexpected pathophysiological consequences. By uncoupling endothelial cell-cell junctions VEGF causes vascular permeability and oedema, resulting in extensive injury to ischaemic tissues after stroke or myocardial infarction. In cancer, VEGF-mediated disruption of the vascular barrier may potentiate tumour cell extravasation, leading to widespread metastatic disease. Therefore, by blocking the vascular permeability promoting effects of VEGF it may be feasible to reduce tissue injury after ischaemic disease and minimize the invasive properties of circulating tumour cells.     

Expression of vascular endothelial growth factor in rat uterus during peri-implantation

The first distinct mark of rodent implantation is the increased vascular permeability and significant angiogenesis at the sites of blastocyst implantation, but its mechanism is not clearly defined. Vascular endothelial growth factor (VEGF) is the key mediator for angiogenesis during embryogenesis and adult span and also serves as a vascular permeability factor. The aim of this study is to explore VEGF regulation mechanism and the possible role that VEGF plays in implantation by studying the VEGF expression and angiogenesis in the rat uterus during estrous cycle, ovarioectomized and peri-implantation stages usingin situ message RNA hybridization and confocal laser scanning techniques. The results indicated that VEGF was regulated by ovarian steroid hormones. VEGF expression before implantation was localized at luminal epithelium, shifted to stroma as implantation initiated and extensively located at the decidualizing stroma region after implantation. Bandeiraea simplicifolia-1 (BS-1) agglutinin and antibody against von Willebrand factor (vWF) were used to mark the endothelial cells and blood vessels. The results showed that the active angiogenesis occurred during the implantation process and this effect was probably mediated by VEGF. The results suggest that under the regulation of ovarian steroid hormones, VEGF plays an essential role in angiogenesis and increasing vascular permeability in endometrium, which are necessary for successful implantation.     

Vascular endothelial growth factor is a potential tumour angiogenesis factor in human gliomas in vivo.

Clinical and experimental studies suggest that angiogenesis is a prerequisite for solid tumour growth. Several growth factors with mitogenic or chemotactic activity for endothelial cells in vitro have been described, but it is not known whether these mediate tumour vascularization in vivo. Glioblastoma, the most common and most malignant brain tumour in humans, is distinguished from astrocytoma by the presence of necroses and vascular proliferations. Here we show that expression of an endothelial cell-specific mitogen, vascular endothelial growth factor (VEGF), is induced in astrocytoma cells but is dramatically upregulated in two apparently different subsets of glioblastoma cells. The high-affinity tyrosine kinase receptor for VEGF, flt, although not expressed in normal brain endothelium, is upregulated in tumour endothelial cells in vivo. These observations strongly support the concept that tumour angiogenesis is regulated by paracrine mechanisms and identify VEGF as a potential tumour angiogenesis factor in vivo.     

红景天苷上调大鼠心肌梗死后心肌组织VEGF的表达

目的:探讨红景天苷对大鼠心肌梗死后心肌组织血管内皮生长因子(VEGF)的表达调控作用及其可能机制。方法:建立大鼠心梗模型后,随机分为模型组、红景天苷3个不同剂量组,每组8只大鼠,另设假手术组8只。术后48 h,红景天苷组分别给予低、中、高,即10,20,40 mg·(kg·d)-1灌胃;模型组和假手术组给予生理盐水20 ml·(kg·d)-1灌胃。4周后处死大鼠,取大鼠心肌组织,反转录PCR(RT-PCR)法测定VEGF mRNA的表达。应用免疫组化和免疫印迹法分析左心室心肌组织VEGF蛋白的表达情况。结果:RT-PCR结果表明,和模型组相比,红景天苷各剂量组VEGF mRNA的表达均明显升高(P<0.01)。免疫组化和免疫印迹分析结果表明,和模型组相比,红景天苷各剂量组心肌组织胞浆中VEGF蛋白的表达均显著升高(P<0.01)。结论:红景天苷通过上调VEGF的表达而促大鼠心肌梗死后心肌组织的血管新生。     

Identification of an angiogenic mitogen selective for endocrine gland endothelium

The known endothelial mitogens stimulate growth of vascular endothelial cells without regard to their tissue of origin. Here we report a growth factor that is expressed largely in one type of tissue and acts selectively on one type of endothelium. This molecule, called endocrine-gland-derived vascular endothelial growth factor (EG-VEGF), induced proliferation, migration and fenestration (the formation of membrane discontinuities) in capillary endothelial cells derived from endocrine glands. However, EG-VEGF had little or no effect on a variety of other endothelial and non-endothelial cell types tested. Similar to VEGF, EG-VEGF possesses a HIF-1 binding site, and its expression is induced by hypoxia. Both EG-VEGF and VEGF resulted in extensive angiogenesis and cyst formation when delivered in the ovary. However, unlike VEGF, EG-VEGF failed to promote angiogenesis in the cornea or skeletal muscle. Expression of human EG-VEGF messenger RNA is restricted to the steroidogenic glands, ovary, testis, adrenal and placenta and is often complementary to the expression of VEGF, suggesting that these molecules function in a coordinated manner. EG-VEGF is an example of a class of highly specific mitogens that act to regulate proliferation and differentiation of the vascular endothelium in a tissue-specific manner.     

Vascular endothelial growth factor induced by hypoxia may mediate hypoxia-initiated angiogenesis.

Inefficient vascular supply and the resultant reduction in tissue oxygen tension often lead to neovascularization in order to satisfy the needs of the tissue. Examples include the compensatory development of collateral blood vessels in ischaemic tissues that are otherwise quiescent for angiogenesis and angiogenesis associated with the healing of hypoxic wounds. But the presumptive hypoxia-induced angiogenic factors that mediate this feedback response have not been identified. Here we show that vascular endothelial growth factor (VEGF; also known as vascular permeability factor) probably functions as a hypoxia-inducible angiogenic factor. VEGF messenger RNA levels are dramatically increased within a few hours of exposing different cell cultures to hypoxia and return to background when normal oxygen supply is resumed. In situ analysis of tumour specimens undergoing neovascularization show that the production of VEGF is specifically induced in a subset of glioblastoma cells distinguished by their immediate proximity to necrotic foci (presumably hypoxic regions) and the clustering of capillaries alongside VEGF-producing cells.     

Effects of vascular endothelial growth factor on angiogenesis of the endothelial cells isolated from cavernous malformations

Human cerebral cavernous malformation （CM） is a common vascular malformation of the central nervous system. We have investigated the biological characteristics of CM endothelial cells and the cellular and molecular mechanisms of CM angiogenesis to offer new insights into exploring effective measures for treatment of this disease. The endothelial cells were isolated from CM tissue masses dissected during operation and expanded in vitro. Expression of VEGFR-1 and VEGFR-2 was examined with immunocytochemical staining. Proliferation, migration and tube formation of CM endothelial cells were determined using MTT, wounding and transmigration assays, and three-dimensional collagen type I gel respectively. The endothelial cells were successfully isolated from the tissue specimens of 25 CMs dissected without dipolar electrocoagulation. The cells show the general characteristics of the vascular endothelial cells. Expression of VEGFR-1 and VEGFR-2 on the cells is higher than that on the normal cerebral microvascular endothelial cells. After treatment with VEGF, numbers of the proliferated and migrated cells, the maximal distance of cell migration and the length and area of capillary-like structures formed in the three-dimensional collagen gel increase significantly. These results demonstrate that expression of VEGFR-1 and VEGFR-2 on CM endothelial cells is up-regulated. By binding to receptors, VEGF may activate the downstream signaling pathways and promote proliferation, migration and tube formation of CM endothelial cells. VEGF/VEGFR signaling pathways play important regulating roles in CM angiogenesis.     

重组人血管内皮生长因子165在大肠杆菌中的可溶性表达、纯化与体外活性评价

人血管内皮生长因子165(VEGF165)可有效促进血管新生和增加血管通透性,在伤口愈合方面有重要医疗价值。建立获取高纯度、高活性的优质重组VEGF165蛋白的方法具有重要意义。研究利用带有6组氨酸标签的二硫键形成蛋白A(Dsb A)的E.coli表达系统实现了Dsb A-VEGF165融合蛋白的可溶性表达;诱导过程中添加5%(v/v)的乙醇可显著提高工程菌中可溶性融合蛋白表达水平。融合蛋白通过Ni亲和层析粗纯,并经牛肠激酶酶切去除标签蛋白。随后利用肝素亲和层析精纯获得重组人VEGF165蛋白。非还原及还原SDS-PAGE电泳检测到分子量为约40 k Da的同源二聚体蛋白,促HUVEC细胞增殖实验显示重组蛋白具有较优的活性,EC50为13 ng/m L。研究实现了Dsb A-VEGF165的在E.coli中可溶性表达,建立了经济、高效的纯化方法,获得了高质量、高活性的重组人VEGF165蛋白。     

血管内皮生长因子对女性生殖功能的调控及其临床意义

血管内皮生长因子是一种特异性血管生长因子，在血管的发生中发挥重要作用。在卵巢、子宫内膜、胎盘等部位有丰富的VEGF及其受体的表达，对女性生殖功能发挥重要的调控作用。近年来VEGF在妇产科领域的临床研究结果表明，VEGF的异常表达与卵巢癌、妊娠高血压、子宫内膜异位症有着密切的关系。     

血管内皮生长因子与急性白血病骨髓的血管新生

目的:探讨血管内皮生长因子(VEGF)在急性白血病及其骨髓的新生血管之间的关系,为白血病的治疗寻找新的治疗方法。方法:查阅总结近15年来国内外相关文献,对VEGF的性质作用特点以及与急性白血病的关系进行综述。结果:白血病细胞表达较高的VEGF,VEGF促使血管生成和内皮细胞增生,白血病细胞与骨髓新生血管之间存在密切的关系。结论:抗VEGF和抗新生血管治疗有可能成为治疗急性白血病的新的思路和方法。