基质金属蛋白酶-2、基质金属蛋白酶-9和血管内皮生长因子在视网膜母细胞瘤中的表达及意义

目的:研究基质金属蛋白酶-2(Matrix metalloproteinase-2,MMP-2)、基质金属蛋白酶-9(Matrix metalloproteinase-9,MMP-9)和血管内皮生长因子(Vascular endothehal growth factor,VEGF)在视网膜母细胞瘤(Retinoblastoma,RB)中的表达及其与RB分化程度和视神经浸润的关系,探讨它们在RB浸润、转移过程中的作用和临床意义.方法:采用免疫组化方法检测40例RB中MMP-2、MMP-9和VEGF的表达.结果:40例RB中MMP-2、MMP-9和VEGF的阳性表达率分别为52.5%、57.5%和72.5%.未分化型RB中MMP-2、MMP-9和VEGF的阳性表达均高于分化型(P＜0.05);有视神经浸润的RB中MMP-2、MMP-9和VEGF的阳性水平均高于无视神经浸润的RB(P＜0.05).RB中MMP-2、MMP-9和VEGF的表达呈正相关关系(P＜0.05).结论:RB中MMP-2、MMP-9和VEGF高表达与RB的浸润和转移相关,且MMP-2、MMP-9表达与VEGF表达存在相关性.联合检测MMP-2、MMP-9和VEGF的表达可作为反映RB浸润、转移潜能的生物学指标,有助于筛选转移高危病例及评价患者预后.     

基质金属蛋白酶-1、血管内皮生长因子在视网膜母细胞瘤中的表达及意义

目的:检测基质金属蛋白酶(Matrix metalloproteinase,MMP)-1及血管内皮生长因子(Vascular endothelial growth factor,VECF)在视网膜母细胞瘤组织中表达情况,分析其与临床分级之间关系的临床意义.方法:采用免疫组化方法检测31例视网膜母细胞瘤组织MMP-1、VEGF表达,对两者的相关性进行统计学分析.结果:MMP-1、VEGF蛋白在视网膜母细胞瘤组织中呈高表达,MMP-1主要表达于肿瘤细胞胞浆或胞膜中,VEGF主要表达于肿瘤细胞胞浆中,MMP-1、VEGF表达和临床分期密切相关(P＜0.05),且其表达在不同临床分期间差异有统计学意义(P＜0.05),MMP-1、VEGF两者表达存在正相关(r=0.787,P＜0.05).结论:MMP-1、VEGF蛋白在视网膜母细胞瘤组织中呈高表达,两者表达与肿瘤临床分级密切相关,可能均参与视网膜母细胞瘤新生血管的生成,在肿瘤的浸润转移等中起重要作用.     

基质金属蛋白酶-2、基质金属蛋白酶-9和血管内皮生长因子在视网膜母细胞瘤中的表达

　　目的　观察基质金属蛋白酶（MMP）-2、MMP-9和血管内皮生长因子(VEGF)在视网膜母细胞瘤(RB)中的表达及其与RB分化程度和视神经浸润的关系。方法　经病理检查确诊的40例RB眼球石蜡标本纳入研究。按照RB细胞中有无菊形团排列以及RB肿瘤细胞是否浸润视神经分为分化型、未分化型以及视神经浸润、视神经无浸润组。其中,分化型15例,未分化型25例;视神经浸润13例,视神经无浸润27例。采用免疫组织化学方法检测其MMP-2、MMP-9和VEGF的表达,对比分析MMP-2、MMP-9和VEGF表达与肿瘤病理组织分型和视神经是否侵润之间的关系。结果　40例RB中,MMP-2、MMP-9和VEGF的阳性表达率分别为52.5%、57.5% 和72.5% 。未分化型RB中MMP-2、MMP-9和VEGF的阳性表达均高于分化型RB中MMP-2、MMP-9和VEGF的阳性表达,差异有统计学意义(χ²=9.037,9.253,8.095;P＜0.05);有视神经浸润的RB中MMP-2、MMP-9和VEGF的阳性水平均高于无视神经浸润RB中MMP-2、MMP-9和VEGF的阳性水平,差异有统计学意义(χ²=11.045,10.243,8.956;P＜0.05)。RB中MMP-2、MMP-9和VEGF的表达呈正相关关系(r=0.126,0.314;P＜0.05)。结论　RB细胞内有MMP-2、MMP-9和VEGF的表达。MMP-2、MMP-9表达与VEGF表达存在相关性。MMP-2、MMP-9和VEGF的表达水平与视神经浸润存在密切的关系。       

基质金属蛋白酶-9、血管内皮钙黏蛋白在翼状胬肉的表达及相关性

目的 研究翼状胬肉组织中基质金属蛋白酶-9(MMP-9)和血管内皮钙黏蛋白(VE-cadherin)的表达,以探讨MMP-9和VE-cadherin表达与翼状胬肉之间的关系,为翼状胬肉的治疗提供理论依据.方法 应用免疫组织化学法检测14例(14只眼)翼状胬肉组织中MMP-9和VE-cadherin的表达,其中6例为复发性翼状胬肉患者,将其与10例正常结膜组织进行比较.结果 翼状胬肉组织中MMP-9和VE-cadherin的表达均高于正常结膜组织,在复发性翼状胬肉组织中的表达明显高于初发性翼状胬肉.另外翼状胬肉组织中MMP-9和VE-cadherin阳性表达间存在正相关(P＜0.05).结论 MMP-9和VE-cadherin的高表达在翼状胬肉的发生和发展中起着重要作用.     

葡萄膜恶性黑色素瘤中血管内皮生长因子水平与肿瘤转移的关系

目的：分析葡萄膜恶性黑色素瘤患者血管内皮细胞生长因子(vascular endothelial growth factor,VEGF)的表达水平是否与葡萄膜恶性黑色素瘤的转移和预后有关。

方法：采用ELISA法检测葡萄膜恶性黑色素瘤患者和正常对照组中外周血VEGF水平,并采用RT-PCR和Western-blot检测葡萄膜恶性黑色素瘤患者肿瘤组织中VEGF mRNA和VEGF蛋白表达水平。对葡萄膜恶性黑色素瘤患者进行随访,用Kaplan-Meier法分析基线VEGF水平与患者术后转移的关系。

结果：葡萄膜恶性黑色素瘤患者外周血VEGF水平较健康者升高。就诊时肿瘤发生转移的患者VEGF mRNA和VEGF蛋白表达水平均较未发生转移的患者升高。在随访过程中,血清VEGF≥700pg/mL的患者,肿瘤发生转移的风险较高。

结论：葡萄膜恶性黑色素瘤患者中VEGF的表达水平与葡萄膜恶性黑色素瘤的转移密切相关。     

实验性脉络膜新生血管中环氧合酶-2、血管内皮生长因子和基质金属蛋白酶-2的表达

目的研究实验性脉络膜新生血管（CNV）大鼠中环氧合酶-2（COX-2）、血管内皮生长因子（VEGF）和基质金属蛋白酶-2（MMP-2）的表达,并进一步探讨其在CNV中的作用机制及相互关系。方法应用氪红激光光凝视网膜的方法诱导制作BN大鼠的CNV模型,分别选取并制作无任何干预的对照组和实验组光凝后第3、7、14、21、30天的＂最大CNV膜＂切片,采用免疫组织化学技术检测COX-2、VEGF和MMP-2在视网膜和CNV膜中的表达。应用HPIAS-1000型高清晰度彩色病理图文分析系统测定COX-2、VEGF和MMP-2阳性染色的平均灰度值。结果 COX-2、VEGF和MMP-2在正常视网膜和脉络膜中呈弱表达,视网膜光凝后,COX-2、VEGF和MMP-2在视网膜和CNV膜中的表达在7～14 d逐渐增强,21 d时三者的表达强度均达到高峰,之后略减弱。视网膜光凝后7、14、21、30 d,实验组COX-2、VEGF和MMP-2的免疫组织化学染色平均灰度值与对照组相比差异均有统计学意义（P〈0.05）。COX-2的表达强度与VEGF和MMP-2的变化均呈正相关（r=0.967,P=0.007;r=0.966,P=0.007）。结论激光诱导的实验性CNV中COX-2、VEGF及MMP-2的表达随着时间的延长呈动态变化,CNV局部炎症诱导的COX-2表达可能是VEGF和MMP-2的上游调节因子。     

组织蛋白酶B和血管内皮生长因子在高氧诱导的视网膜新生血管中的作用

背景 血管内皮生长因子(VEGF)在血管发育和新生血管形成中起着关键作用.研究表明,组织蛋白酶B(Cathepsin B)参与新生血管的生成,但其具体机制尚不明确. 目的 探讨Cathepsin B和VEGF在高氧诱导小鼠视网膜新生血管中的表达及二者之间的关系. 方法 应用随机数字表法将44只7日龄清洁级C57BL/6J小鼠随机分为正常对照组、高氧诱导组、空白对照(NC)-绿色荧光蛋白(GFP)-慢病毒(Lv)组和Cathepsin B-RNA干扰(RNAi)-Lv组,每组11只小鼠22只眼.正常对照组小鼠在自然环境中生长;其余3个组7日龄小鼠置于氧体积分数为(75±2)％的密闭氧箱内饲养5d,之后返回到正常环境中.高氧诱导组的12日龄小鼠不给予任何药物干预;NC-GFP-Lv组和Cathepsin B-RNAi-Lv组的12日龄小鼠分别给予玻璃体腔注射NC-GFP-Lv和Cathepsin B-RNAi-Lv各1μl.取各组17日龄小鼠,颈椎脱臼法处死后剥取视网膜,分别采用real-time PCR和Western blot法检测小鼠视网膜中Cathepsin B和VEGF mRNA及蛋白的相对表达量.结果 荧光显微镜下Cathepsin B-RNAi-Lv组视网膜新生血管分层和分支均较高氧诱导组和NC-GFP-Lv组少.各组Cathepsin B和VEGF mRNA总体比较,差异均有统计学意义(F=444.89,P=0.00;F=519.78,P=0.00),其中高氧诱导组、NC-GFP-Lv组和Cathepsin B-RNAi-Lv组小鼠视网膜中Cathepsin B和VEGFmRNA的相对表达量均明显高于正常对照组,Cathepsin B-RNAi-Lv组小鼠视网膜中Cathepsin B和VEGF mRNA的相对表达量均明显低于高氧诱导组和NC-GFP-Lv组,差异均有统计学意义(均P＜0.05).各组Cathepsin B和VEGF蛋白总体比较,差异均有统计学意义(F=54.37,P=0.00;F=79.65,P=0.00),其中高氧诱导组、NC-GFP-Lv组和Cathepsin B-RNAi-Lv组小鼠视网膜中Cathepsin B和VEGF蛋白的相对表达水平均明显高于正常对照组,Cathepsin B-RNAi-Lv组小鼠视网膜中Cathepsin B和VEGF蛋白的相对表达水平均明显低于高氧诱导组和NC-GFP-Lv组,差异均有统计学意义(均P＜0.05). 结论 高氧诱导的视网膜新生血管中Cathepsin B和VEGF高表达,Cathepsin B-RNAi-Lv可以抑制Cathepsin B和VEGF mRNA和蛋白的表达水平,VEGF表达的改变可能是受Cathepsin B表达的影响.     

葡萄膜黑色素瘤患者血清血管内皮生长因子浓度及临床意义

目的测定葡萄膜黑色素瘤患者血清血管内皮生长因子(VEGF)的水平,并探讨其临床意义。方法应用酶联免疫吸附法(ELISA),分别检测不同组织病理分型的葡萄膜黑色素瘤(27例)、葡萄膜黑色素细胞瘤患者(6例)以及正常人(16例)外周血清中VEGF的质量浓度。结果正常成人和葡萄膜黑色素细胞瘤患者血清VEGF浓度显著低于葡萄膜黑色素瘤患者;有巩膜导水管浸润和远处转移的葡萄膜黑色素瘤患者血清VEGF质量浓度明显高于不伴巩膜导水管浸润和远处转移的患者;梭形细胞型患者血清VEGF浓度与混合型和上皮细胞型患者经统计分析无显著差异。结论血清VEGF水平在监测葡萄膜黑色素瘤的转移中有一定的意义。     

血管生成素-1对糖尿病大鼠视网膜微血管病变、基质金属蛋白酶和血管内皮生长因子的抑制作用

目的 观察血管生成素-1(angiogenin-1,Ang-1)对糖尿病大鼠视网膜微血管痛变和基质金属蛋白酶( matrix metalloproteinases,MMP)、血管内皮生长因子(vascular endothelial growth factor,VEGF)表达的抑制作用.方法 通过尾静脉注射链脲佐茵素( streptozotocin,STZ)建立糖尿病大鼠模型.随机分配治疗组大鼠20只,每15 d每眼球后注射Ang-1溶液0.05mL 1次(4ng·L-1),糖尿病阳性对照组20只不给予任何处理.2组与正常对照组(20只)分别饲养3个月、6个月时,采用眼底荧光素血管造影(fundus fluorescoin angiography,FFA)观察视网膜血管改变;免疫组化法定量检测各组大鼠视网膜VEGF的表达;蛋白印迹法定量检测MMP的表达量.结果 FFA观察结果:3个月时糖尿病阳性对照组大鼠视网膜毛细血管扭曲、不规则,Ang-1治疗组可见明显的改善;6个月时,糖尿病阳性对照组可见视网膜微动脉瘤,Ang-1治疗组与糖尿病阳性对照组相比微动脉瘤减少.免疫组化检测结果显示,3个月、6个月时正常对照组仅见微弱的VEGF表达,灰度值分别为92.968 7±2.0400和81.0311±2.7700,糖尿病阳性对照组表达明显上调,灰度值分别为120.0008 ±2.9011和124.790 5±1.9615,与正常对照组比较差异均具有显著统计学意义(P＜0.05、P＜0.01);Ang-1治疗组3个月、6个月时VEGF表达灰度值分别为116.154 8±1.343 1和92.850 9±2.323 4,与糖尿病阳性对照组相比差异有统计学意义(P＜0.05).蛋白印迹法定量检测结果显示,正常对照组MMP蛋白无表达,3个月时Ang-1治疗组表达量高于糖尿病阳性对照组(P＜0.05),6个月时糖尿病阳性对照组与Ang-1治疗组间差异无统计学意义(P＞0.05).结论 Ang-1能有效改善糖尿病大鼠视网膜的微血管病变,并抑制视网膜内VEGF、MMP的表达.     

眼部新生血管与抗血管内皮生长因子治疗

眼部新生血管性病变是致盲的主要原因之一,可见于多种眼底疾病,其确切的发病机制尚不完全清楚.大量的研究证据表明血管内皮生长因子(VEGF)是新生血管形成的关键调控因子.本文对VEGF的特性、VEGF在眼部新生血管形成中的作用和抗VEGF治疗进行综述.     

血管内皮生长因子拮抗剂治疗眼内新生血管性疾病研究进展

血管内皮生长因子(vascular endothelia growth factor,VEGF),是迄今为止鉴定出来最重要的血管生成因子,与新生血管形成和通透性紧密相关。眼内新生血管形成以及与其相伴的渗出、出血、纤维结缔组织增生等一系列病变严重破坏眼的结构和功能,是引起患者视力下降乃至失明的重要原因。近年来,把VEGF作为靶点来治疗眼内新生血管和/或血管通透性增加的疾病已取得突破性进展,多种相关药物已进入临床试验阶段,在短期内显示出良好的安全性和有效性,然而,也应注意到该类治疗潜在的并发症,还应进一步研究以确定其长期的安全性和治疗效果。     

明胶酶A及血管内皮细胞生长因子在翼状胬肉组织中的表达

目的探讨血管内皮细胞生长因子(vascularendothelialgrowthfactor,VEGF)、明胶酶A与翼状胬肉发生、浸润的关系。方法术中切除的31例翼状胬肉标本制成冰冻切片,经免疫组织化学方法检测翼状胬肉组织中VEGF、明胶酶A的表达,并与正常结膜组织中VEGF、明胶酶A的表达对比,以半定量分法评价并统计其结果。结果正常结膜组织与翼状胬肉上皮组织VEGF表达阴性、弱阳性、强阳性例数分别为6、2、0和0、8、23(χ2=29.187),明胶酶A表达阴性、弱阳性、强阳性例数分别为8、0、0和0、8、23(χ2=39·00),差异都有极显著意义(P<0.001)。静止期与进展期翼状胬肉上皮组织VEGF表达弱阳性、强阳性例数分别为4、19和4、4(χ2=4.216,P<0.05),明胶酶A表达弱阳性、强阳性例数分别为2、21和6、2(χ2=12.816,P<0.001),差异有显著意义。结论VEGF、明胶酶A在翼状胬肉组织中过度表达,其表达量与翼状胬肉的病理分期密切相关,提示二者在翼状胬肉的形成和浸润过程中起重要作用。     

血管内皮生长因子与眼部新生血管性疾病

血管内皮生长因子（ ＶＥＧＦ） 是一种广泛分布于大多数器官和组织血管内皮细胞的多肽类因子,它是迄今为止所发现的惟一作用于血管内皮的生长因子,可诱导血管生成。综述了ＶＥＧＦ 的生物学特性与眼部新生血管性疾病的研究进展     

Matrix metalloproteinase-9 and vascular endothelial growth factor expression change in experimental retinal neovascularization

AIM: To investigate the signal transduction mechanism of matrix metalloproteinase-9 (MMP-9) mediated- vascular endothelial growth factor (VEGF) expression and retinal neovascularization (RNV) in oxygen-induced retinopathy (OIR) model. METHODS: C57BL/6J mice were divided into four groups: control group, OIR group, OIR control group (phosphate-buffered saline by intravitreal injection) and treated group [tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) by intravitreal injection]. OIR model was established in C57BL/6J mice exposed to 75%±2% oxygen for 5d. mRNA level and protein expression of MMP-9, TIMP-1 and VEGF were measured by real-time polymerase chain reaction and Western blotting, and located by immunohistochemistry. RESULTS: Levels of MMP-9 and VEGF in retina were significantly increased in animals with OIR and OIR control group. Levels of TIMP-1 in retina was significantly reduced in animals with OIR and OIR control group. Furthermore, a significant correlation was found between MMP-9 and VEGF. Intravitreal injection of TIMP-1 significantly reduced MMP-9 and VEGF expression of the OIR mouse model (all P<0.05). CONCLUSION: These results demonstrate that MMP-9-mediated up-regulation of VEGF promotes RNV in retinopathy of prematurity (ROP). TIMP-1 may be a potential target for the prevention and treatment of ROP.     

Expressions of survivin and vascular endothelial growth factor in a Murine model of proliferative retinopathy

AIM: To examine the expression of survivin and vascular endothelial growth factor(VEGF) during the development of retinal neovascularization (NV) in a mouse model. METHODS: A well-characterized murine model of retinal NV was used to study the expression of survivin and VEGF. NV of the retina was induced in mice by exposure to 75% O2 from postnatal day P7 to P12, followed by return to room air from P12 to P17. Expression of survivin and VEGF protein was analyzed by Immunohistochemistry. In addition, mouse model of proliferative retinopathy was analyzed by retinal fluorescein angiography and quantification analysis. RESULTS: The normal mice had both superfiekal and deep vascular layers that extended from the optic nerve to the periphery. In intraocular pressure(IOP) mice were characterized by represent a typical pattern of pathological retinal NV. There are less or little nuclei of new vessels vascular endothelial cell breaking through the inner retinal than in retinopathy of prematurity (ROP) mice, large clusters of blood vessels were adherent to the internal limiting membrane(ILM) (0.27±0.20 vs 23.38±1.027, t=9.454, P<0.001). During the angiogenic period from P13 to P17, survivin and VEGF protein expression increased in experimental retinas compared with control samples(2.56±0.46 vs 3.34±0.40, t=17.43, P<0.01: 2.18±0.75 vs 4.34±0.25, t=19.61, P<0.01). Protein levels of VEGF and survivn has significantly positive correlation (P<0.05, r=0.411). CONCLUSION: Correlation was made at the protein levels of survivin expression compared with that of VEGF in a murine model of retinal NV, which suggests a temporal role for survivin and VEGF in new vessel formation in response to hypoxic stimulation.     

Expressions of survivin and vascular endothelial growth factor in a Murine model of proliferative retinopathy

AIM: To examine the expression of survivin and vascular endothelial growth factor(VEGF) during the development of retinal neovascularization (NV) in a mouse model. METHODS: A well-characterized murine model of retinal NV was used to study the expression of survivin and VEGF. NV of the retina was induced in mice by exposure to 75% O2 from postnatal day P7 to P12, followed by return to room air from P12 to P17. Expression of survivin and VEGF protein was analyzed by Immunohistochemistry. In addition, mouse model of proliferative retinopathy was analyzed by retinal fluorescein angiography and quantification analysis. RESULTS: The normal mice had both superfiekal and deep vascular layers that extended from the optic nerve to the periphery. In intraocular pressure(IOP) mice were characterized by represent a typical pattern of pathological retinal NV. There are less or little nuclei of new vessels vascular endothelial cell breaking through the inner retinal than in retinopathy of prematurity (ROP) mice, large clusters of blood vessels were adherent to the internal limiting membrane(ILM) (0.27±0.20 vs 23.38±1.027, t=9.454, P<0.001). During the angiogenic period from P13 to P17, survivin and VEGF protein expression increased in experimental retinas compared with control samples(2.56±0.46 vs 3.34±0.40, t=17.43, P<0.01: 2.18±0.75 vs 4.34±0.25, t=19.61, P<0.01). Protein levels of VEGF and survivn has significantly positive correlation (P<0.05, r=0.411). CONCLUSION: Correlation was made at the protein levels of survivin expression compared with that of VEGF in a murine model of retinal NV, which suggests a temporal role for survivin and VEGF in new vessel formation in response to hypoxic stimulation.     

血管内皮生长因子 A的选择性剪接与眼内新生血管的生成

眼内新生血管的生成是多种眼病致盲的重要原因.血管内皮生长因子A(VEGF-A)家族是促进眼内新生血管生成的关键因素,其通过调控病理性血管的发生和增加血管的通透性而起作用.VEGF-A依选择性剪接方式的不同,可形成2个蛋白家族,分别是具有促血管生成作用的VEGFxxx家族和具有抗血管生成作用的VEGFxxxb家族.VEGFxxxb家族蛋白在正常眼组织中均有表达,而在糖尿病性视网膜病变患者的眼组织中表达水平降低.VEGF165b是VEGFxxxb家族中最早分离出来且研究最为广泛的分子结构,其可以明显抑制视网膜前新生血管的生成,而对视网膜生理性血管的发生无抑制作用.随着对VEGFxxxb家族研究的逐步深入,选择性剪接调节VEGFxxx与VEGFxxxb两者之间的平衡,可作为糖尿病性视网膜病变、年龄相关性黄斑变性等眼内新生血管性疾病的治疗新策略.

Abstract：

Ocular neovascularization is the primary cause of blindness in a wide range of ocular diseases. The vascular endothelial growth factor A (VEGF-A) is the key factor involved in ocular angiogenesis, which can cause eye diseases through the development of pathological angiogenesis and increase of vascular permeability. There are two families of VEGF-A isoforms formed by alternative splicing,the angiogenic VEGF-A family ( VEGFxxx ), known to contribute to ocular neovascularization, and the antiangiogenic VEGF-A family ( VEGFxxx b), which is found in normal ocular tissues but downregulated in human diabetic retinopathy. The first member of the VEGFxxxb family to be isolated was VEGF165b. It can significantly reduce preretinal neovascularization without inhibition of physiological intraretinal angiogenesis.As the studies on the VEGFxxxb family proceed more deeply, controlling the balance of VEGFxxx to VEGFxxxb isoforms may be therapeutically valuable in the treatment of angiogenic eye diseases such as diabetic retinopathy and age-related macular degeneration.