多次玻璃体腔注射抗血管内皮生长药物对黄斑水肿患者角膜生物力学的作用

目的观察多次玻璃体腔注射抗血管内皮生长因子(VEGF)药物对角膜生物力学的作用。方法前瞻性病例对照研究。选取玻璃体腔注射抗VEGF治疗的患者共64例(64只眼),其中,湿性老年性黄斑变性(nAMD)19例(19只眼),糖尿病黄斑水肿(DME)20例(20只眼),视网膜静脉阻塞(RVO)黄斑水肿25例(25只眼)。阿柏西普(2 mg/0.05 ml)治疗29例(29只眼),雷株单抗(0.5 mg/0.05 ml)治疗35例(35只眼)。经过3次连续注射治疗的患者共64例(64只眼),经过4次连续注射治疗的患者共10例(10只眼)。使用眼反应分析仪(ORA)对角膜生物力学指标进行测量。比较每次注药后1个月与注射前的角膜阻力因子(CRF)、角膜滞后量(CH)、角膜补偿眼压值(IOPcc)、可重复的模拟Goldmann眼压值(IOPg)的变化。比较使用阿柏西普和雷株单抗两种不同药物多次玻璃体腔注射后以上所有指标的变化,同时比较nAMD、DME、RVO三种不同疾病多次玻璃体腔注射抗VEGF药物后以上所有指标的变化。结果(1)雷株单抗/阿柏西普多次注射后角膜生物力学各指标在术后各时间点较术前相比,差异无统计学意义;(2)注射次数与角膜生物力学各项指标的改变无明显相关性;(3)术前基线时,角膜生物力学等各指标在3种不同疾病患者基线时差异无统计学意义;(4)注射3次后,DME组的CH值较RVO组和AMD组升高,且差异有统计学意义。结论多次玻璃体腔注射抗VEGF药物使DME组的CH值则显著高于RVO组和AMD组。糖尿病黄斑水肿患者多次玻璃体腔注射抗VEGF药物对角膜生物力学会产生一定影响。

血管内皮生长因子-C在肾癌中的表达及意义

目的:探讨血管内皮生长因子-C(VEGF-C)在肾癌发生发展中的作用。方法:应用免疫组织化学SP法检测VEGF-C在肾癌及正常肾组织中的表达情况。结果:肾癌组织VEGF-C的阳性表达率(85.0%)显著高于正常肾组织(P<0.01)。肾癌组织VEGF-C的阳性表达与年龄、性别、分化程度无关(P>0.05),与有无淋巴结转移和肾癌临床分期有关(P<0.05)。结论:VEGF-C可能通过促进淋巴管生成参与肾癌的淋巴结转移,在肾癌的发生发展中发挥重要作用。

血管内皮生长因子-B与眼部新生血管性疾病

异常的新生血管会导致多种眼部疾病,而血管内皮生长因子（VEGF）在新生血管的发生及发展中起重要作用.VEGF-B作为VEGF家族的一员,对血管生成和血管通透性无显著影响,但可通过血管生存作用和细胞凋亡作用对新生血管的生长进行调控,从而抑制新生血管的发生和发展.同时,VEGF-B对心脏和神经元等具有保护作用.抗VEGF治疗作为目前明确有效的治疗新生血管的方法,一直受到广泛关注,而因VEGF-B的两面性,对于抗VEGF-B的药物在治疗眼科新生血管性疾病方面的作用尚需进一步研究.就VEGF-B对机体血管、细胞、神经元及心脏所起的作用及其在眼部新生血管性疾病治疗中的应用进行综述.

血管内皮细胞VEGF在鼠尾再植术后的表达及加味活血祛瘀糖浆干预的实验研究

目的:探讨大鼠鼠尾离断再植术后血管内皮细胞VEGF的表达及加味活血祛瘀糖浆对再植术后VEGF表达的影响。方法:采用大鼠鼠尾离断再植造模,ELISE分别检测对照组、实验组及药物组血清中VEGF的表达;免疫组化法检测3组实验大鼠血管组织中VEGF的表达。结果:ELISE、免疫组织化学染色均证实鼠尾再植后血清和组织中VEGF表达升高,VEGF免疫组化检测后8小时两者已有显著表达,VEGF在24小时表达达高峰,VEGF表达持续约5天,血清ELISA检测同样显示VEGF表达较对照组明显升高。应用加味活血祛瘀糖浆能使血清和组织中的VEGF明显升高,促进血管修复。结论:大鼠鼠尾离断后血清中VEGF表达增加,加味活血化瘀糖浆组中再植鼠尾后血中VEGF的表达增加,加味活血化瘀糖浆可通过促VEGF表达上调,内皮细胞增殖,从而保护血管内皮细胞,利于离断血管的修复。

抗血管内皮生长因子类药物在新生血管性青光眼及滤过手术抗瘢痕治疗中的应用

新生血管性青光眼（NVG）是一种由多种疾病导致视网膜缺血缺氧而发生的继发性青光眼,研究发现其病因与VEGF有关。目前,抗VEGF药物作为治疗NVG的手段之一,已成为现阶段青光眼研究领域的热点,为提高NVG疗效开辟了新的途径。本文就有关NVG病理生理基础与临床过程,包括Bevacizumab、Ranibizumab、Pegaptanib及Aflibercept在内的4种抗VEGF药物的基础研究,抗VEGF药物在NVG及滤过手术抗瘢痕治疗中应用的国内外研究进展进行综述,以期为临床治疗NVG、提高手术成功率提供参考。

血管内皮生长因子在糖尿病肾病中的作用及其机制

血管内皮生长因子在糖尿病肾病中起重要作用,高血糖引起的蛋白激酶C、氧化应激、缺血缺氧等都能诱导血管内皮生长因子的过表达.大量研究表明血管内皮生长因子可通过多种途径调控肾小球各类细胞及基质的结构和功能,参与糖尿病肾病的病理生理过程,并在疾病进展中起重要作用.这些发现为糖尿病肾病的治疗提供了新的思路.

关注湿性年龄相关性黄斑变性抗血管内皮生长因子治疗中的无应答病例

抗VEGF药物已经成为湿性AMD患者的一线标准用药，但是仍有部分患者治疗后出现无应答，其原因目前仍不确切，但可能与AMD的类型、黄斑区解剖因素异常、遗传基因型的不同、AMD病变的程度及抗药反应有关。眼科医师应当关注这部分无应答病例并给予正确的处理，最大限度地避免抗VEGF药物的滥用，并积极探索适合患者最佳个性化治疗措施。（中华眼科杂志，2014，50：406-410）

子宫内膜癌组织中缺氧诱导因子-1α、血管内皮生长因子基因表达的临床意义

目的 探讨HIF-1α、VEGF基因表达与子宫内膜癌临床分期间关系.方法 对30例子宫内膜癌患者的癌组织应用免疫组织化学方法进行HlF-1α、VEGF的检测,分析不同的病理分型和临床分期的表达情况.另取同期存档的子宫内膜不典型增生组织蜡块10例作为对照.结果 H1F-1α、VEGF在子宫内膜不典型增生及子宫内膜癌各期的阳性表达率差异均有统计学意义（x2=11.87、8.71,均P＜0.05）;超声肿瘤血流分级与HIF-α、VEGF间存在相关性.病变与HIF-1α、VEGF间存在相关性.结论 HIF-α、VEGF表达程度与子宫内膜癌恶性程度呈正相关.HIF-1α、VEGF子宫内膜癌的预后和转移关系密切.

人表皮生长因子受体2在结直肠肿瘤组织中的表达及其与血管内皮生长因子A的相关性

目的：研究人表皮生长因子受体2（human epidermal receptor 2，HER-2）在结直肠癌中的表达及其与血管内皮生长因子A（vascular endothelial growth factor A，VEGFA）的相关性。方法选择结直肠癌患者80例为结直肠癌组，另以距肿瘤组织＞5 cm的并经病理学证实的癌旁正常结肠组织30例作为对照组，采用免疫组织化学染色SP法，对结直肠癌组及对照组组织中的HER-2、VEGF-A、FⅧ的表达水平给予检测，并探讨HER-2与VEGF-A、微血管密度（MVD）的相关性。结果结直肠癌组的结肠癌组织中的HER-2表达阳性率为61．25％，明显高于对照组癌旁正常结直肠组织中HER-2表达阳性率的3．33％（χ2＝6.342，P＜0．05）。结直肠癌组的结肠癌组织中的VEGF-A表达阳性率为65．00％，明显高于对照组癌旁正常结肠组织中VEGF-A表达阳性率的3．33％（χ2＝4．725，P＜0．05）。结直肠癌患者癌组织中HER-2、VEGF-A表达均与肿瘤分化程度、淋巴结转移、临床分级相关（P＜0．05），与性别及肿瘤大小无关（P＞0.05）。经Pearman等级相关分析可知，结直肠癌组织中HER-2的表达与VEGF-A表达呈正相关（r＝0．736，P＜0．05）。HER-2阳性表达病例MVD值为（61．5±9．4），对照组MVD值为（48．3±7．8），两组比较差异有统计学意义（t＝8．440，P＜0．01）。结论 HER-2的表达在结直肠癌的侵袭与转移中具有十分重要的作用，推测结直肠癌组织中的HER-2高表达可能为通过促进VEGF-A在体内的表达而增加患者血管密度，从而加速结直肠癌肿瘤的发生及进展。

电脉冲介导的血管内皮细胞生长因子基因治疗对大鼠随意型皮瓣成活的影响

目的探讨血管内皮细胞生长因子(vascularendothelialgrowthfactor,VEGF)质粒直接皮下注射,结合直流电脉冲刺激的基因治疗方案的血管生成效应及不同剂量质粒对皮瓣成活的影响。方法设计大鼠随意皮瓣模型,利用直流电脉冲将皮下注射的重组质粒PCMVMCSVEGFIRES2EGFP导入缺血皮瓣内。观察皮瓣成活的情况,利用苏木素伊红(HE)染色,观察VEGF基因的血管生成效应。结果皮下直接注射结合直流电脉冲成功地将重组质粒导入皮肤。基因转染后7d观察到明显的血管增生。基因治疗组皮瓣成活面积[80μg为(77.38±4.56)%,400μg为(82.57±5.21)%]显著大于对照组[(48.96±4.32)%]。但两基因治疗组皮瓣成活面积差异无统计学意义。结论VEGF质粒直接皮下注射结合直流电脉冲刺激能够高效转染大鼠皮肤,具有明显的血管生成效应,能够显著促进大鼠随意型皮瓣的成活。质粒的剂量对皮瓣成活的影响差异无统计学意义。

外源性表达VEGF165b对人膀胱癌裸鼠移植瘤生长影响的研究

目的建立人膀胱癌T24细胞裸鼠移植瘤模型,探讨外源性表达VEGF165b对其生长的影响及其机制。方法分别移植未转染和转染VEGF165b表达载体的人膀胱癌T24细胞到裸鼠膀胱内,28d使处死动物,称量肿瘤重量,测定肿瘤体积。Western blot法检测肿瘤VEGF165b、p-VEGFR2、AKT、p-AKT蛋白表达。结果转染pcDNA-VEGF165b组肿瘤重量和体积与未转染组和转染pcDNA3.0组相比明显降低(P<0.05),而未转染组和转染pcDNA3.0组比较无统计学差异(P>0.05)。Western blot结果显示,转染pcDNA-VEGF165b组与未转染组和转染pcDNA3.0组比较,VEGF165b表达增高,p-VEGFR2、p-AKT表达降低,而未转染组和转染pcDNA3.0组间无明显差异。结论外源性表达VEGF165b可通过抑制VEGFR2信号通路传导而明显抑制人膀胱癌裸鼠移植瘤生长。

Effect of acetyl L-carnitine on human retinal pigment epithelium-19 cells in hypoxic conditions

AIM:To investigate the effect of acetyl-L-carnitine(ALCAR)on cell viability,morphological integrity,and vascular endothelial growth factor(VEGF)expression in human retinal pigment epithelium(ARPE-19)cells using a hypoxic model.METHODS:In the first set of experiments,the optimal CoCl_(2) dose was determined by exposing ARPE-19 cell cultures to different concentrations.To evaluate the effect of ALCAR on cell viability,five groups of ARPE-19 cell culture were established that included a control group,a sham group(200μM CoCl_(2)),and groups that received 1,10 and 100 mM doses of ALCAR combined with 200μM CoCl_(2),respectively.The cell viability was measured by MTT assay.The morphological characteristics of cells were observed by an inverted phase contrast microscope.The levels of VEGF and HIF-1α secretion by ARPE-19 cells were detected by enzyme linked immunosorbent assay(ELISA)assay.RESULTS:ARPE-19 cells were exposed to different doses of CoCl_(2) in order to create a hypoxia model.Nevertheless,when exposed to a concentration of 200μM CoCl_(2),a notable decrease in viability to 83% was noted.ALCAR was found to increase the cell viability at 1 mM and 10 mM concentrations,while the highest concentration(100 mM)did not have an added effect.The cell viability was found to be significantly higher in the groups treated with a concentration of 1 mM and 10 mM ALCAR compared to the Sham group(P=0.041,P=0.019,respectively).The cell viability and morphology remained unaffected by the greatest dose of ALCAR(100 mM).The administration of 10 mM ALCAR demonstrated a statistically significant reduction in the levels of VEGF and HIF-1α compared with the Sham group(P=0.013,P=0.033,respectively).CONCLUSION:The findings from the current study indicate that ALCAR could represent a viable therapeutic option with the potential to open up novel treatment pathways for retinal diseases,particular relevance for age-related macular degeneration(AMD).However,to fully elucidate ALCAR’s application potential in retinal diseases,additional investigation is necessary to clearly define the exact mechanisms involved.

Hydroxysafflor Yellow A Promotes Vascular Endothelial Cell Proliferation via VEGF/VEGF Receptor

Aim To study the proliferative effeet of hydroxysaftlor yellow A （HSYA） on cultured canine aortic endothelial cell （VEC） in normoxic （21% O2 ） or hypoxic （10% O2 ） culture and the underlying mechanism. Methods The endothelial cells were scratched from trypsined canine aorta endothelium. HSYA was added to the cells at final concentrations of 1 × 10^-3, 1 × 10^-4 and 1 × 10^-5 mol· L^-1, respectively. VEGF （2.6 × 10^-7 mol· L^-1 ）-treated cells were used as the positive control. The proliferative effect of HSYA on VEC was determined at 48, 72, 96, and 120 h in normoxic culture by MTI＂ assay. Similarly, the proliferation of VEC was determined at 12, 24, 48, and 72 h in hypoxic culture by MTF assay. The effects of HSYA on VEC proliferation and VEGF secretion were investigated by MTr and ELISA assays at the presence of the antibodies to VEGF and VEGF receptors. Results Pretreatment with HSYA at concentrations of 1 × 10^-3 and 1 × 10^-4 mol· L^-1 enhanced VEC proliferation in normoxic culture. The most significant enhancing effect of HSYA on VEC proliferation was achieved at 24, 48, and 72 h in hypoxic culture in concentration-dependent and time-dependent manner. HSYA at 1 × 10^-3 mol·L^-1 showed a potency similar to VEGF at 2.6 × 10^-7 mol·L^-1 . Pretreatment with the antibodies of Flt-1, KDR or VEGF blocked the proliferative effect of HSYA with similar potencies. Antibodies of Fit-1 or VEGF antagonized the promoting effect of HSYA on VEGF secretion. Conclusion HSYA promotes VEC proliferation either in normoxic or hypoxic culture, especially in the latter condition. This effect of HSYA is at least partly mediated by VEGF and VEGF receptor.

Real-time Quantitative PCR Analysis of Vascular Endothelial Growth Factor Receptor-2(VEGFR-2) Expression at Zebrafish Different Developmental Stages

[Objective]To investigate the expression of zebrafish vascular endothelial growth factor-2（VEGFR-2） at different developmental stages.[Method]Total RNAs were extracted from 12,24,48,72 and 96 hpf stage zebrafish embryos and larvae.Real-time quantitative RT-PCR was performed to examine the expression of VEGFR-2.The data were analyzed by 2^-△△Ct method.[Result]The expression level of VEGFR-2 gene increased gradually from 12 to 72 hpf,and subsequently decreased at 96 hpf.The expression level was lowest at 12 hpf,highest at 72 hpf,and had significant differences when compared with that of other developmental stages.[Conclusion]The expression level of VEGFR-2 increases gradually before blood vessel maturation and decreases as blood vessels mature.

IMPACTS ON THE EXPRESSION OF VASCULAR ENDOTHELIAL GROWTH FACTOR IN ISCHEMIC BRAIN TISSUE TREATED WITH CLUSTER PUNC-TURE ON SCALP ACUPOINTS IN RATS

Objective To probe into the impacts on the expression of vascular endothelial growth factor （VEGF） in ischemic brain tissue treated with cluster puncture on scalp acupoints in rats, Methods 128 rats were randomized into pseudo-operation group, model group, scalp-needling group and cluster-needling group, In scalp-needling group, penetration method was used on the focal side from Bǎihuì （百会 GV20） to Qǔbīn （曲鬓 GB7）, and in cluster-needling group, penetration method was used on both sides from GV20 to GB7, and a common needling on GV 20. Needles were punctured 2 mm in depth, constantly rotated for 10min, retained for 2 h. Immunohistrochemical method was applied to determine VEGF expression and microvessel density （MVD）. Results With the intervention of cluster puncture on scalp acupoints, VEGF： expressions on every time-spot after ischemia were enhanced apparently, superior to those in scalp-needling group. On the three time-spots of the 7^th, 14^th and 21^st days, MVD was increased after cluster puncture on scale acupoints, superior to those in scalp-needling group. Conclusion Cluster puncture on scalp acupoints up-regulated VEGF expression and promoted regeneration of microvessel.

COX-2和VEGF-C与胃癌淋巴结转移的相关性研究

目的研究环氧化酶-2(COX-2)与血管内皮生长因子-C(VEGF-C)在胃癌中的表达与生物学行为的关系。方法采用免疫组织化学染色法,检测54例胃癌原发灶标本COX-2与VEGF-C蛋白表达的情况,分析其与胃癌淋巴转移之间的关系。结果所测54例胃癌组织中,COX-2及VEGF-C的表达阳性率分别为70·4%(38/54)和66·7%(36/54)。COX-2及VEGF-C表达与淋巴结转移呈明显相关性(P<0·01),但与肿瘤分化程度和有无浆膜浸润等无明显相关(P>0·05)。VEGF-C与肿瘤的TNM分期相关(P<0·01)。结论COX-2及VEGF-C在胃癌组织中的表达与肿瘤的淋巴转移密切相关;COX-2可能通过上调VEGF-C表达,促进胃癌淋巴管生成,并顺次导致淋巴结转移的发生。

Expression of Angiopoietin-2 and Vascular Endothelial Growth Factor in Oral Squamous Cell Carcinoma and Its Significance

Objective： To investigate the expression of angiopoietin-2 （Ang-2） and vascular endothelialcell growth factor （VEGF） in oral squamous cell carcinoma （OSCC） and their correlations with clinicopathologic parameters, angiogenesis and vessel maturation of OSCC. Methods： The expression of Ang-2 and VEGF was detected in 41 speciments of human OSCC, 30 adjacent noncancerous oral tissues and 10 specimens of normal oral mucosa by conventional immumohistochemistry. Microvessel density （MVD） and vessel maturation index （VMI） were also assessed by double-labelling immumohistochemistry staining against CD34, a marker of pan-endothelial cells, and that against alpha-smooth muscle actin （α-SMA）, a marker of mural cells （pericytes/smooth muscle cells）. Results： The positive expression rate of Ang-2 and VEGF in 41 OSCC tissues was 51.22% and 63.42%, respectively. The expression of Ang-2 and VEGF was significantly higher in OSCC than in adjacent noncancerous oral tissues （all P〈0.05） and normal oral mucosa （all P〈0.05）. In the clinicopathologic parameters, the Ang-2 expression was closely correlated with tumor lymph node metastasis （P〈0.01） and the VEGF expression was correlated with tumor differentiated degree （P〈0.05）, but there was no significant correlation among the Ang-2 and VEGF expression and patients＇ sex, age and TNM stages （all P〉0.05）. The MVD of OSCC positive for both Ang-2 and VEGF was significantly higher than that of OSCC negative for both Ang-2 and VEGF （P〈0.05）. The VMI of OSCC positive for Ang-2 was significantly lower than that of OSCC negative for Ang-2 （P〈0.05）. When Ang-2 expression was combined with the staus of VEGF expression, MVD of OSCC positive for both Ang-2 and VEGF was the highest （51.08±2.99） as compared with that of other status in patient with OSCC （all P〈0.05）. Conclusion： The overexpression of Ang-2 and VEGF may play a crucial role in the development of OSCC. They are closely associated with angiogenesis and vessel maturation of tumor.

Increased protein and mRNA expression of endostatin in the ischemic brain tissue of rabbits after middle cerebral artery occlusion

Objective To explore the changes of endostatin （a strong anti-angiogenesis factor） and vascular endothelial growth factor （VEGF） in the brain tissues of rabbits following cerebral ischemia induced by middle cerebral artery occlusion （MCAO）. Methods Twenty-four New Zealand white rabbits were randomly divided into 5 groups： control （n = 5）, sham-operation （n = 4）, 2-hour ischemia （n = 5）, 24-hour ischemia （n = 5）, and 48-hour ischemia （n = 5）. The expression of VEGF and endostatin were measured by enzyme-linked immunosorbent assay （ELISA） and immunohistochemistry, respectively. In situ hybridization was used to characterize the expression of mRNA for the endostatin. Results Both the protein （at least 50%, P 〈 0.01） and mRNA （at least 70%, P 〈 0.05） of endostatin increased significantly in the ischemic brain tissues after MCAO compared with the control group. VEGF increased at least 270% in the brain after cerebral ischemia （P 〈 0.05）. Conclusion Cerebral ischemia leads to an up-regulation of endostatin in the brain, which is not associated with the increase of VEGF in the brain. The increase of endostatin may serve as a deleterious mechanism for ischemic injury through blocking angiogenesis.

Effects of Ghrelin Antisense Inhibition on VEGF and Its Receptor Flt-1 mRNA Expression

[ Objective] This study was to investigate the effect of VEGF and its receptor Fit-1 mRNA expression in Mongolia sheep umbilical vein endothelial cells by ghrelin antisense inhibition. [ Method] Experiments were divided into 4 groups： group Ⅰ （blank control group） ; group Ⅱ （liposome group） ; group Ⅲ （SCON group： 20 μmol/L sense oligonucleotide） ; group Ⅳ （ASCON： 20 μmol/L antisense oligonucleotide）. VEGF and its receptor Fit-1 mRNA expression changes were detected by using real-time fluorescence quantitative detection after 24, 36 and 48 h. [ Result] The expression of VEGF mRNA in group Ⅰ, group Ⅱ were insignificantly different at higher expression levels, and did not change significantly with the time; the expression of VEGF mRNA in group Ⅲ assumed a slight decrease, but there were no significant differences between group I and group Ⅱ （P 〉0.05）, the expression of VEGF mRNA in group Ⅳ（antisense oligonucleotide group ） decreased significantly （P 〈 0.05） ; the expression of VEGF receptor FLT-1 mRNA was similar to that of VEGF. [ Conclusion] Antisense inhibition ghrelin has a downward effect to the expression of VEGF and its receptor Fit-1 the mRNA.

Vascular Endothelial Growth Factor C Expression in Gastric Carcinoma and Its Relationship with Lymph Node Metastasis

Objective: To study the expression of vascular endothelial growth factor C (VEGF-C) in gastric carcinoma and its relationship with lymph node metastasis. Methods: The expression of VEGF-C mRNA in 5 gastric carcinoma cell lines was detected by reverse transcription-polymerase chain reaction (RT-PCR). Simultaneously, the expression of VEGF-C protein in gastric carcinoma tissues, which were obtained from 63 patients who underwent radical gastrectomy, was detected by immunohistochemistry. Results: Three of the 5 gastric carcinoma cell lines, MKN-45, SGC-7901 and AGS, expressed VEGF-C mRNA. VEGF-C protein was expressed in 52.4% (33/63) of patients. VEGF-C protein expression was more frequently found in tumors with lymph node metastasis than in those without (P<0.01). VEGF-C protein expression was also closely related to lymphatic invasion (P<0.01) and TNM stage (P<0.01). However, there was no significant correlation between VEGF-C expression and the age, gender, tumor size, tumor location, Lauren classification, depth of invasion, and vascular invasion. Conclusion: The expression of VEGF-C is closely related to lymph node metastasis of gastric carcinoma, and lymphangiogenesis might be a new target for treatment of gastric carcinoma.