新生血管性青光眼患者房水中血小板源性生长因子-C和血管内皮生长因子水平的测定和分析

背景 新生血管性青光眼(NVG)是由不同病因引起的严重致盲眼病,与视网膜组织缺氧后分泌因子有关,如血管内皮生长因子(VEGF)等,但仅用抗VEGF疗法并不能完全抑制新生血管的生长.我们先前的研究发现,血小板源性生长因子-C(PDGF-C)参与眼内病理性新生血管的生成,但PDGF-C在NVG中的作用尚不清楚. 目的 定量检测NVG患者房水中VEGF和PDGF-C的质量浓度,为NVG的治疗提供新的思路. 方法 采用前瞻性队列研究方法,收集2014年1月至2015年8月在第四军医大学西京医院就诊的NVG患者62例62眼,其中10眼近1年内曾行视网膜光凝和/或冷凝治疗,其他52眼中原发病为视网膜中央静脉阻塞(CRVO)者16眼,糖尿病视网膜病变(DR)者20眼,视网膜脱离(RD)术后者5眼,视网膜血管炎(Eales病)者4眼,未知原因者7眼;虹膜新生血管Ⅱ级者13眼,Ⅲ级者29眼,Ⅳ级者10眼.同期纳入年龄相关性白内障患者11例11眼作为对照.分别于手术中采集受检眼房水0.1 ～0.2 ml,应用ELISA法检测受检眼房水中VEGF和PDGF-C质量浓度. 结果 NVG组患眼房水中VEGF和PDGF-C质量浓度分别为(1 138.17±69.31)ng/L和(29.80±1.64) ng/L,明显高于对照组的(679.54±49.81)ng/L和(18.60± 1.85) ng/L,差异均有统计学意义(t=20.95、20.49,均P＜0.01).视网膜光凝和/或冷凝组患眼房水中VEGF和PDGF-C质量浓度分别为(1 095.99±52.71) ng/L和(28.55±0.94) ng/L,均低于非光凝和/或冷凝组的(1 146.28±69.57) ng/L和(30.04±1.64) ng/L,差异均有统计学意义(t=-2.160,P=0.034;t=-2.760,P=0.008).NVG患者房水中VEGF与PDGF-C质量浓度呈正相关(r=0.346,P=0.006).不同原发病组间及不同虹膜新生血管分级组间患眼房水中VEGF和PDGF-C质量浓度的总体比较差异均无统计学意义(均P＞0.05). 结论 NVG患者房水中VEGF和PDGF-C质量浓度明显升高,视网膜光凝和/或冷凝治疗可抑制VEGF和PDGF-C的产生,靶向抑制PDGF-C可为NVG的抗新生血管治疗提供新的选择.     

新生血管性青光眼转化生长因子β2含量及其相关影响因素

目的 观察新生血管性青光眼(NVG)眼内房水和玻璃体液中转化生长因子(TGF)-β2总量和活化含量,并分析其相关影响因素.方法 前瞻性研究NVG患者54例(54只眼)房水和玻璃体液中总TGF-β2(tTGF-β2)和活化TGF-β2 (aTGF-β2)含量,其中视网膜中央静脉阻塞(CRVO)17只眼,糖尿病视网膜病变(...     

新生血管性青光眼患者眼内血管内皮生长因子浓度对虹膜新生血管的影响

目的　观测手术前后新生血管性青光眼 (neovascular glaucoma,NVG)患者房水及玻璃体中血管内皮生长因子 (vascular endothelial growth factor,VEGF)含量变化 ,及对 NVG虹膜新生血管的影响。方法　应用酶联免疫吸附测定法 (enzyme linked im munosorbent assay,EL ISA)对 18例 NVG患者内眼手术前后取得的 72份房水、玻璃体标本进行 VEGF检测对比 ,观察手术前后虹膜新生血管的消退情况。结果　 NVG患者手术后房水中VEGF含量为 (0 .5 76± 0 .196) ng/ml,较手术前房水 VEGF含量 (1.486± 0 .3 2 2 ) ng/ml明显降低 (t=10 .3 94,P<0 .0 0 1) ,而玻璃体中的 VEGF含量由 (1.619± 0 .3 84) ng/ml降为 (0 .682± 0 .2 2 2 ) ng/m l(t=9.3 93 ,P <0 .0 0 1) ,两组间差别均有非常显著性意义。术后发现两周内 77.78% (14/18)患者虹膜新生血管消退。结论　 NVG患者虹膜新生血管形成及维持可能依赖于房水、玻璃体中 VEGF含量 ,手术后眼内 VEGF浓度下降可能促进虹膜新生血管的消退     

前部视网膜冷凝术对新生血管性青光眼患者房水中血管内皮生长因子含量的影响

目的 探讨前部视网膜冷凝术对新生血管性青光眼患者房水中血管内皮生长因子（VEGF）含量的影响以及VEGF含量的变化与虹膜新生血管之间的关系。方法对28例确诊为新生血管性青光眼患者行虹膜血管造影,确定新生血管的范围和数量后,行前部视网膜冷凝术,7～14d后经虹膜血管造影确定虹膜新生血管大部分消退后,再行小梁切除术。分别于前部视网膜冷凝术前和小梁切除术前抽取房水标本,另取30例老年性白内障患者房水标本。采用酶联免疫吸附法分别测定全部房水标本中的VEGF含量。结果小梁切除术前房水中VEGF的含量[（2．096±0．512）ng／ml]明显低于前视网膜冷凝术前房水中VEGF含量[（0．478±0．312）ng／ml],两者比较差异有统计学意义（P〈0．01）。小梁切除术前房水中VEGF含量明显高于老年性白内障患者房水中VEGF的含量[（0．198±0．045）ng／ml],两者比较差异有统计学意义（P〈0．01）。结论VEGF在虹膜新生血管的形成中可能发挥一定的作用;阻断促使虹膜产生新生血管的VEGF来源,可抑制新生血管性青光眼的发生.（中华眼科杂志．2007．43：622-625）     

新生血管性青光眼患者眼内液血管 内皮生长因子的检测

目的了解新生血管性青光眼(neovascular glaucoma, NVG)患者房水及玻璃体中血管内皮生长因子(vascular endothelial growth factor, VEGF)的含量以及对虹膜新生血管发生的影响。方法应用酶联免疫吸附测定法(enzyme linked immu nosorbent assay, ELISA)分别对11例NVG患者行内眼手术时抽取的房水、玻璃体22个标本以及同期6例黄斑裂孔手术患者(对照组)房水、玻璃体12个标本进行VEGF检测。结果NVG患者房水、玻璃体中VEGF含量分别为(1.451±0.247)、(1.610±0.125) ng/ml,较对照组房水、玻璃体中的 VEGF含量(0.189±0.038)、(0.201±0.055) ng/ml明显增高, 两组间差别有非常显著性意义(t=12.007,P<0.001;t=26.057,P<0.001)。结论NVG患者房水、玻璃体中VEGF浓度显著增高,提示VEGF在NVG患者虹膜新生血管形成过程中可能具有一定作用。(中华眼底病杂志,2001,17:305-306)     

肝细胞生长因子在糖尿病新生血管性青光眼玻璃体内含量的对比研究

目的研究糖尿病新生血管性青光眼(NVG)患者玻璃体内肝细胞生长因子(HGF)的含量改变及其与血管内皮生长因子(VEGF)的相关性。设计前瞻性病例系列。研究对象糖尿病NVG患者14例(14眼),不伴虹膜新生血管的增生性糖尿病视网膜病变(PDR)患者22例(22眼),特发性黄斑裂孔(IMH)患者19例(19眼)。方法采用双抗体夹心酶联免疫吸附分析法(ELISA)检测三种疾病(三组)患者玻璃体内HGF及VEGF浓度改变。主要指标同一标本内HGF和VEGF浓度。结果NVG组、PDR组及IMH组的玻璃体内HGF浓度分别为(12908．42±2946．46)、(9770．86±3802．99)、(4160．54±2044．80)pg/ml(Kruskal-Walls检验,X~2=32．36,P=0．000),HGF浓度组间两两比较,差异均有统计学意义(P均＜0．01);三组的玻璃体内VEGF浓度分别为(823．50±718．58)、(821．82±786．27)、(22．73±3．20)pg/ml(X~2=28．30,P=0．000)。各组同一标本的HGF浓度与VEGF浓度之间无相关性(P均＞0．05)。结论糖尿病NVG玻璃体内HGF浓度明显高于不伴虹膜新生血管的PDR及LMH者,提示HGF可能参与NVG发病过程中视网膜及虹膜新生血管的形成过程。但其玻璃体内HGF与VEGF浓度水平无相关性。(眼科,2007,16:40-43)     

增生性玻璃体视网膜疾病眼内液血管内皮生长因子的表达

目的 观察增生性玻璃体视网膜疾病患者眼内液（房水、玻璃体）中血管内皮生长因子（VEGF）含量的变化规律，探讨VEGF在增生性玻璃体视网膜疾病发展变化中的作用。 方法 采用双抗体夹心酶联免疫吸附试验(ELISA)定量检测增生性玻璃体视网膜病变（PVR）、视网膜静脉阻塞（RVO）、增生性糖尿病视网膜病变（PDR）、新生血管性青光眼（NVG）患者组及正常对照组房水、玻璃体VEGF含量。 结果 PVR、RVO、PDR、NVG患者组房水及玻璃体VEGF含量均高于正常对照组，其差异均有统计学意义(P<0.05)；NVG、PDR、RVO、PVR患者组房水及玻璃体VEGF含量依次降低 ，其差异有统计学意义(P<0.05)；PVR、RVO、PDR、NVG患者组和正常对照组玻璃体VEGF含量均高于房水VEGF含量，其差异有统计学意义(P<0.05)；PVR患者病史与房水、玻璃体VEGF含量呈负相关（r分别为－0.819、－0.823，P＜0.05）；RVO患者病史与房水、玻璃体VEGF含量呈正相关（r分别为0.913、0.929，P＜0.05）；PDR患者玻璃体积血时间与房水、玻璃体VEGF含量呈正相关（r分别为0.905、0.920，P＜0.05）。 结论 增生性玻璃体视网膜疾病患者房水及玻璃体VEGF含量明显增高，VEGF可能在增生性玻璃体视网膜疾病发展变化中起着重要的作用。 (中华眼底病杂志， 2006， 22：313-316）     

新生血管性青光眼患者房水中血小板反应蛋白－1的表达及其与血管内皮生长因子相关关系的临床研究

目的检测新生血管性青光眼(NVG)与年龄相关性白内障患者房水中血小板反应蛋白－1(TSP－1)的表达情况,并分析NVG患者房水中的TSP－1与血管内皮生长因子(VEGF)的关系。 方法纳入30例(30只眼)NVG患者(NVG患者组)以及30例(30只眼)年龄相关性白内障患者(对照组)。所有NVG患者在行抗青光眼手术前抽取房水,同时在白内障手术时抽取年龄相关性白内障患者房水。采用酶联免疫吸附试验检测房水以及血液中TSP－1与VEGF的浓度。房水和血液中VEGF浓度和TSP－1浓度采用均数±标准差( ±s)表示,采用独立样本t检验进行组间比较。各细胞因子之间、细胞因子浓度与年龄、眼压之间的相关性用采用Spearman's相关分析。 结果NVG组患者房水中TSP－1浓度为(15.47±2.28) ng/ml,而对照组患者房水中TSP－1浓度为(2.28±1.59)ng/ml,NVG组患者房水中TSP－1浓度明显高于对照组,差异有统计学意义(t＝12.70,P<0.05)。NVG组患者与对照组患者房水中VEGF浓度分别为(3355.81±2284.54)pg/ml及(262.68±125.66 )pg/ml,差异有统计学意义(t＝7.41,P<0.05)。NVG组患者血浆中TSP－1浓度为(0.65±0.27)ng/ml,对照组患者血浆中TSP－1浓度为(0.57±0.25)ng/ml,两组之间浓度相当,差异无统计学意义(t＝1.16,P>0.05)。NVG组患者的VEGF血浆浓度为(13.01±7.60)pg/ml,对照组患者的VEGF血浆浓度为(14.62±6.96)pg/ml。二者比较,差异无统计学意义(t＝－0.86,P>0.05)。NVG患者房水中TSP－1与VEGF浓度呈显著正相关(r＝0.479,P<0.05)。在NVG组中,不管是TSP－1因子还是VEGF,其房水中的浓度与血浆中的浓度均无显著相关性。同样,在对照组中,房水TSP－1浓度与血浆TSP－1浓度无相关性,且房水VEGF浓度与血浆VEGF浓度亦无相关性。 结论NVG患者房水中TSP－1与VEGF表达明显上调,明显上调的TSP－1与VEGF呈现显著正相关。     

血管内皮生长因子干扰RNA对鼠视网膜中血管内皮生长因子mRNA表达的抑制作用

目的 研究血管内皮生长因子（vascular endothelial cell growth factor,VEGF）小片段干扰RNA（small interference RNA,siRNA）对鼠视网膜VEGF mRNA的抑制作用,探讨其对视网膜新生血管治疗的可行性.方法 体外培养人鼻咽癌细胞（CNE-2Z）,分成正常氧培养组（20% O2）和低氧培养组（1% O2）.采用脂质体（LF 2000）将VEGF siRNA转染两组细胞,RT-PCR检测VEGF mRNA的表达,确立VEGF siRNA对VEGFmRNA的抑制效率.然后,建立高浓度氧（75%）诱导的C57BL./6J小鼠视网膜新生血管动物模型,以脂质体为载体,将VEGF siRNA重组质粒注射到鼠玻璃体腔内,RT-PCR检测视网膜组织中VEGF mRNA的表达水平.结果 正常氧培养的CNE-2Z细胞有VEGF mRNA表达,低氧状态下VEGFmRNA表达增多,两者之间差异有显著性（P＜0.01）;与未转染组和转染空载体组相比,在正常氧和低氧状态下,VEGFsiRNA均能明显抑制VEGFmRNA的表达（P＜0.01）;正常氧状态下VEGF siRNA的抑制效率比低氧状态高.高浓度氧诱导的C57BL/6J小鼠视网膜新生血管动物模型中,玻璃体腔注射VEGF siRNA组视网膜组织中VEGF mRNA表达明显下降（P＜0.01）.结论 VEGF特异的siRNA能有效地抑制人鼻咽癌细胞CNE-2Z和C57BL/6J 小鼠视网膜新生血管动物模型视网膜中VEGF mRNA的表达.     

血清及房水中VEGF、IL-6水平与新生血管性青光眼的相关性研究

目的测定新生血管性青光眼（NVG）患者血清及房水中血管内皮生长因子（VEGF）、白细胞介素-6（IL-6）的水平,探讨VEGF、IL-6在NVG发生发展中的作用。方法选取NVG患者20例（20只眼）作为实验组（A组）,原发性慢性闭角型青光眼患者（B组）、老年性白内障患者（C组）各20例（20只眼）作为对照组。采集3组患者血清及房水标本,通过双抗体夹心酶联免疫吸附试验（ELISA法）分别检测血清与房水中VEGF、IL-6的水平。结果 （1）A组房水中VEGF、IL-6的水平（1336.80±70.15）pg/ml、（691.15±50.09）pg/ml明显高于B组（311.60±31.06）pg/ml、（168.25±11.95）pg/ml和C组（165.75±13.95）pg/ml,（92.10±9.59）pg/ml,3组间两两比较,差异均具有统计学意义（F=4019.334,P〈0.01;F=2275.019,P〈0.01）。A组血清中VEGF、IL-6的水平（545.40±155.49）pg/ml、（291.35±22.66）pg/ml高于B组（321.15±52.57）pg/ml、（104.35±13.21）pg/ml和C组（176.30±20.38）pg/ml、（87.00±12.70）pg/ml,3组间两两比较,差异均具有统计学意义（F=75.940,P〈0.01;F=906.947,P〈0.01）。（2）A组房水中VEGF与IL-6的水平呈显著的正相关性,差异具有统计学意义（r=0.857,P〈0.01）。其余各组标本中无显著的相关性（P〉0.05）。结论 NVG患者房水及血清中VEGF、IL-6的水平明显高于对照组,且房水中二者水平呈明显正相关,提示在NVG病理机制过程中,VEGF、IL-6作为促血管生成因子,相互促进、相互影响,共同导致了NVG的发生和发展。     

抗VEGF药物联合PRP治疗新生血管性青光眼的效果

目的：探究抗血管内皮生长因子(vascular endothelial growth factor,VEGF)联合视网膜激光光凝(PRP)治疗新生血管性青光眼(neovascular glaucoma,NVG)的效果,以及对房水中VEGF、血小板源性生长因子-C(platelet derived growth factor-C,PDGF-C)的影响。

方法：选择2016-11/2017-11在我院接受检查和治疗的NVG患者90例93眼,随机分为对照组和观察组。对照组使用PRP治疗,观察组使用PRP联合玻璃体注射雷珠单抗治疗。比较两组患者治疗后1mo的临床疗效、虹膜新生血管和视力恢复情况。并分析治疗前和治疗后1mo视网膜静脉循环时间、眼压、视网膜神经纤维层(retinal nerve fiber layer,RNFL)厚度、视野缺损值、房水中VEGF和PDGF-C水平以及不良反应。

结果：治疗后1mo观察组的临床疗效、虹膜新生血管消失情况和视力恢复情况优于对照组,差异有统计学意义(P<0.05)。治疗后1mo观察组的视网膜静脉循环时间、眼压、视野缺损值、房水VEGF和PDGF-C水平均显著低于对照组,RNFL厚度显著高于对照组,差异有统计学意义(P<0.05)。两组患者不良反应发生情况比较,差异无统计学意义(P>0.05)。

结论：对NVG患者使用视网膜激光光凝联合抗VEGF药物治疗可更有效抑制血管生成,恢复视网膜功能,具有更好的疗效,这可能与联合治疗具有更佳的下调VEGF与PDGF-C的作用有关。     

Increased interleukin-6 in aqueous humor of neovascular glaucoma.

PURPOSE: To demonstrate the involvement of proinflammatory cytokines in intraocular neovascularization by detecting the presence of interleukin (IL)-6, IL-2, and tumor necrosis factor (TNF)-alpha in aqueous humor and serum of patients with neovascular glaucoma (NVG) secondary to central retinal vein occlusion (CRVO). METHODS: According to the grade of iris neovascularization (NVI), patients with CRVO were divided into three groups: CRVO without NVI, CRVO with NVI, and CRVO with regressed NVI. Healthy patients with cataract were enrolled as control subjects. Enzyme-linked immunosorbent assay was used to quantitate the concentrations of the cytokines IL-6, IL-2, and TNF-alpha in aqueous humor and serum from patients with NVG and control subjects. RESULTS: In serum, the levels of IL-6, IL-2, and TNF-alpha did not differ among groups. In aqueous humor, only IL-6 showed significant change among groups. IL-6 levels in aqueous humor of group 2, CRVO with NVI (1532.0+/-221.1 pg/ml; P < 0.001), and group 3, CRVO with regressed NVI (234+/-154.6 pg/ml; P < 0.001), were significantly higher. There was no significant difference in IL-6 levels between the control group (26.4+/-21.8 pg/ml) and group 1 (15.6+/-0.9 pg/ml). CONCLUSIONS: The inflammatory cytokine IL-6 in aqueous humor increased spatially and temporally correlated with the grade of NVI in patients of NVG secondary to CRVO. The aqueous IL-6 increased in NVI and decreased after vessels regressed. It is possible that the significantly higher level of IL-6 was due to intraocular synthesis because of the minimal change in serum. The increased level of IL-6 may have a putative role along with other angiogenic factors in angiogenesis of NVG as a possible predictor of NVI.     

糖尿病视网膜病变患者玻璃体中肝细胞生长因子的含量检测

目的 定量研究肝细胞生长因子(hepatocyte growth factor, HGF)在糖尿病视网膜病变(diabetic retinopathy, DR)患者玻璃体中的水平，探讨HGF在增生性糖尿病视网膜病变 (proliferative diabetic retinopathy, PDR)等新生血管形成病理过程中的作用。 方法 采用双夹心酶联免疫吸附测定法检测对照组10只眼玻璃体以及单纯型DR组7只眼、PDR组33只眼和其它与新生血管生成有关的视网膜疾病组8只眼玻璃体切割手术中所取玻璃体内HGF的含量。PDR组中无虹膜新生血管者24只眼，伴虹膜新生血管者9只眼。 结果 玻璃体中HGF的含量对照组为（3.34±1.9）μg/L；单纯型DR组为（4.8±2.5）μg/L；PDR组中不伴虹膜新生血管生成者为（13.0±5.2）μg/L；PDR伴有虹膜新生血管生成者为（18.6±7.2）μg/L，其它与新生血管生成有关的视网膜疾病组为（12.1±8.9）μg/L。PDR组和其它与新生血管生成有关的视网膜疾病组玻璃体中HGF的含量比对照组显著升高(t=6.49, 5.70, 3.01, P<0.01)；PDR组中伴有虹膜新生血管生成者较PDR不伴虹膜新生血管生成者以及单纯型DR组玻璃体中HGF的含量均高，其差异有显著性意义(t=2.47, P<0.05或t=4.84, P<0.01)。 结论 在PDR和与新生血管生成有关的视网膜疾病患眼玻璃体内HGF的含量升高，提示HGF可能在视网膜新生血管生成的病理过程中起一定作用。 (中华眼底病杂志, 2002, 18: 131-13)     

Clinical factors related to the aqueous levels of vascular endothelial growth factor and hepatocyte growth factor in proliferative diabetic retinopathy

PURPOSE: To investigate clinical factors related to the aqueous humor levels of vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF) in patients with proliferative diabetic retinopathy (PDR). METHODS: Undiluted aqueous humor was obtained during ocular surgery from 46 eyes of 46 patients and the levels of growth factors were measured using enzyme-linked immunosorbent assays. VEGF and HGF levels were compared with the number of photocoagulation. VEGF and HGF levels in patient groups stratified according to the existence of vitreous hemorrhage, retinal detachment, and fibrovascular membrane were analyzed. And clinical parameters associated with the growth factors were determined by multiple regression analysis. RESULTS: The levels of VEGF decreased significantly as the extent of photocoagulation increased and showed significant positive correlation with the existence of vitreous hemorrhage. There was no significant correlation between VEGF levels and the existence of fibrovascular membrane and traction retinal detachment. The levels of HGF also decreased significantly as the extent of photocoagulation increased, but increased significantly when fibrovascular membrane existed. There were no significant correlations between HGF levels and the existence of vitreous hemorrhage and traction retinal detachment. Concerning systemic conditions, each growth factor has no significant correlation with duration and type of diabetes mellitus, the treatment regimen, the control of hemoglobin A1C, and the existence of hypertension or renal dysfunction. CONCLUSION: The therapeutic effect of panretinal photocoagulation on PDR might be partly exerted by reduction of the levels of VEGF and HGF in ocular fluid. Since the clinical parameters associated with VEGF were different from those associated with HGF, these growth factors might influence the progression of retinopathy in different ways.     

Concentration of Vascular Endothelial Growth Factor After Intracameral Bevacizumab Injection in Eyes With Neovascular Glaucoma

Purpose

To study the concentration of vascular endothelial growth factor (VEGF) in the aqueous humor before and after intracameral injection of bevacizumab in eyes with neovascular glaucoma, and to detect the duration of an anti-VEGF effect of bevacizumab in the anterior chamber.

Methods

In this prospective interventional case series, 1.25 mg of bevacizumab was injected into the anterior chamber of five eyes in five neovascular glaucoma patients. Aqueous humor samples were obtained just before intracameral injection of bevacizumab and two weeks after injection. The concentrations of VEGF in the aqueous humor were measured using ELISA. To investigate corneal endothelial damage after intrecameral bevacizumab injection, specular microscopy was performed before injection and two weeks after injection. Slit lamp photo and iris fluorescent angiography was performed to determine the regression of iris neovascularization.

Results

After injection, substantial regression of neovascularization or fluorescein leakage was seen in all treated eyes. The VEGF concentrations in the aqueous humor in eyes with NVG were 1181.8±1248.3 pg/mL before intracameral injection of bevacizumab. Two weeks after injection, the VEGF concentrations decreased to 33.2±12.2 pg/mL (p=0.04, Wilcoxon signed rank test). There were no significant changes in IOP or corneal endothelial cells.

Conclusions

Intracameral bevacizumab injection can remarkably reduce iris neovascularization in neovascular glaucoma patients. VEGF levels were significantly decreased two weeks after injection and corneal toxicity was not observed during short term follow-up.