增殖性玻璃体视网膜病变玻璃体血管内皮生长因子表达的研究

目的　研究血管内皮生长因子 (VEGF)在增殖性玻璃体视网膜病变 (PVR)玻璃体中的表达 ,分析VEGF在 PVR中的作用。方法　增殖性玻璃体视网膜病变 37例 ,其中视网膜脱离 PVR2 2例 2 2眼 ,外伤性 PVR15例。以 12例猝死无眼疾的成年人作为正常对照。检测病例组、对照组玻璃体 VEGF水平。VEGF的测定采用对抗夹心法 (EL ISA)。结果　 37例增殖性玻璃体视网膜病变玻璃体 VEGF平均 2 2 0 .5 2± 10 1.97pg/ m l,其中 2 2例网脱PVR玻璃体 VEGF2 38.90± 6 1.2 4pg/ ml,外伤性 PVR玻璃体 VEGF193.5 8± 5 8.17pg/ ml。正常对照组玻璃体VEGF89.90± 36 .36 pg/ ml。PVR玻璃体 VEGF水平明显高于正常对照组 (P <0 .0 5 )。VEGF在 PVRA级、B级水平较高 ,且随 PVR程度增加而降低。有危险因素 PVR,VEGF水平高于无危险因素 PVR(P <0 .0 5 )。结论　作为细胞间的传导信号 ,VEGF参与 PVR的发生发展。VEGF在 PVR中呈上调性表达。VEGF不仅在缺血性眼部疾病中呈高表达 ,而且在某些非缺血性眼部疾病中表达升高。 VEGF主要在 PVR早期发挥作用 ,PVR危险因素可以刺激玻璃体 VEGF上行性表达。     

Unbalanced vitreous levels of pigment epithelium-derived factor and vascular endothelial growth factor in diabetic retinopathy

PURPOSE: To determine the levels of pigment epithelium-derived factor (PEDF) and vascular endothelial growth factor (VEGF) in the vitreous of patients with diabetic retinopathy (DR). DESIGN: Experimental study of PEDF and VEGF levels in vitreous samples collected during vitrectomy. METHODS: The levels of PEDF and VEGF were measured by enzyme-linked immunosorbent assay in the vitreous of 46 eyes of 43 patients who underwent vitrectomy with diabetic retinopathy (DR) (32 eyes of 29 patients) and an idiopathic macular hole (MH) (14 eyes of 14 patients). RESULTS: The vitreal concentration of PEDF was significantly lower at 1.11 +/- 0.14 microg/ml (mean +/- standard error) in eyes with DR than in eyes with MH at 1.71 +/- 0.22 microg/ml (P =.021). The VEGF level was 1799 +/- 478 pg/ml in eyes with DR and not detectable in MH. The PEDF level in proliferative DR (PDR) (0.94 +/- 0.12 microg/ml) was lower than that in nonproliferative DR (NPDR) (2.25 +/- 0.32 microg/ml), and that in active DR (0.85 +/- 0.14 microg/ml) was significantly lower than that in inactive DR (1.59 +/- 0.24 microg/ml; P =.01). The VEGF level was 2025 +/- 533 pg/ml in PDR and 215 +/- 201 pg/ml in NPDR and that in active DR (2543 +/- 673 pg/ml) was significantly higher than that in inactive DR (395 +/- 188 pg/ml; P =.0098). CONCLUSIONS: These results suggest that lower levels of PEDF and higher levels of VEGF may be related to the angiogenesis in DR that leads to active PDR.     

Risk evaluation of outcome of vitreous surgery for proliferative diabetic retinopathy based on vitreous level of vascular endothelial growth factor and angiotensin II

AIMS: To ascertain whether measurement of the vitreous fluid levels of vascular endothelial growth factor (VEGF) or angiotensin II (Ang II) could predict the outcome of vitreous surgery in patients with proliferative diabetic retinopathy (PDR). METHODS: A prospective observational case study was performed in 61 consecutive patients (61 eyes) with PDR who underwent vitreoretinal surgery. Vitreous fluid samples were obtained during surgery. The VEGF level in vitreous fluid and plasma was determined by enzyme linked immunosorbent assay, while the Ang II level was measured by radioimmunoassay. Patients were prospectively followed for 6 months and the postoperative outcome was analysed by logistic regression analysis. RESULTS: No improvement and/or progression of PDR was seen in 15 (25%) of the 61 eyes. Vitreous levels of VEGF and Ang II were significantly higher in eyes with progression of PDR than in eyes with regression of PDR (p = 0.0044, and p = 0.0178, respectively). Multivariate logistic regression analysis showed that the vitreous VEGF level increased along with the progression of PDR after vitreous surgery (odds ratio 2.48, p = 0.0008). CONCLUSION: A high vitreous fluid VEGF level is associated with a significant risk of postoperative progression of PDR. The vitreous level of VEGF at the time of surgery may be a useful predictor of the outcome.     

CFH基因多态性与玻璃体液VEGF对年龄相关性黄斑变性患者抗VEGF疗效影响的交互作用

目的 探讨补体因子H(CFH)基因多态性与玻璃体液血管内皮生长因子(VEGF)对年龄相关性黄斑变性(AMD)患者抗VEGF疗效影响的交互作用。方法 选取2020年5月～2022年1月广州医科大学附属第三医院眼科149例(149眼)AMD患者为研究对象,根据抗VEGF治疗是否有应答,分为应答组(111例)、无应答组(38例)。比较两组患者CFH基因多态性、玻璃体液VEGF表达水平,比较CFH位点rs1061170不同基因型患者玻璃体液VEGF表达水平,采用Logistic分析抗VEGF疗效的相关影响因素,采用交互作用系数γ分析CFH基因多态性、玻璃体液VEGF的交互作用是否存在及其作用类型。结果 无应答组患者CFH位点rs1061170基因型分布与应答组比较,差异有统计学意义(P<0.05)。无应答组患者玻璃体液VEGF高于应答组(P<0.05)。在全部AMD患者中,玻璃体液VEGF高水平患者CFH位点rs1061170基因型分布与低水平患者比较,差异有统计学意义(P<0.05)。在全部AMD患者中,玻璃体液VEGF、CFH位点rs1061170 CC基因型均是抗VEG...     

放射性视网膜病变大鼠玻璃体与血清血管内皮生长因子的相关研究

目的通过放射性^60Co照射SD大鼠制作放射性视网膜病变（RR）模型,探讨在RR的发展中,血管内皮生长因子（VEGF）的作用。方法45只SD大鼠,随机分为正常对照组、10Gy组和30Gy组,每组15只。^60Co射线照射制作RR模型。分别于3、6、9个月时检测RR大鼠血清及玻璃体中的VEGF质量浓度,并制作大鼠视网膜毛细血管铺片。结果30Gy组与10Gy组大鼠玻璃体中VEGF浓度较正常对照组增高,差异均有统计学意义（P〈0.01）。30Gy组与10Gy组大鼠血清中VEGF质量浓度与正常对照组比较差异均无统计学意义（P〉0.05）。实验组大鼠视网膜毛细血管铺片结果显示,随时间延长视网膜微血管病变逐渐加重。结论在RR大鼠模型中,VEGF参与RR的发展。随时间延长玻璃体中VEGF质量浓度增加,后者与RR视网膜微血管病变的发展呈正相关,为临床上治疗RR提供理论依据。     

Intravitreal vascular endothelial growth factor

Purpose

To evaluate whether a specific pre-analytical stabilization regimen is needed for naïve vitreous taps to detect true values of intrinsic VEGF levels.

Methods

Fourteen consecutive patients with different vitreomacular pathologies without blood–retina-barrier breakdown were scheduled for standard 23-gauge three-port pars plana vitrectomy, and naïve vitreous taps were sampled at the beginning of each procedure. The extracted vitreous specimen was split; one half was immediately stored in a ?20 °C freezer (unstabilized samples) and the other half was instantly stabilized with albumin (2.5 % final conc.), followed by arginine stabilization (1.25 M final conc.) and consecutively stored in a ?20 °C freezer (stabilized samples).

Results

Intravitreal VEGF was detected in all 14 analyzed samples (100 %). VEGF levels were shown to be 46.5 pg/ml?±?62.3 pg/ml (MV ± SD; range: 5.99–232.3 pg/ml) in unstabilized, and 120.4 pg/ml?±?94.4 pg/ml (range: 42.9 pg/ml–289.6 pg/ml) in stabilized vitreous samples. Intravitreal VEGF levels in stabilized vitreous samples were on average 2.6-fold, and thus significantly higher than in unstabilized taps of same eyes (p?=?0.001, Wilcoxon test). VEGF levels in stabilized vitreous samples can be up to 8.5 times higher than in corresponding unstabilized vitreous taps of same eyes (bootstrap analysis). Intravitreal VEGF levels in unstabilized samples correlate with those in stabilized vitreous taps (r?=?0.594; p?=?0.025; Pearson).

Conclusions

An adequate pre-analytic stabilization regimen is needed to evaluate the most accurate intravitreal VEGF levels. This in turn will result in a better understanding of the physiological as well as pathological role of VEGF within the eye. Furthermore, knowing the true value of intravitreal VEGF levels will help to calculate the dosage of intravitrealy applied anti-VEGF drugs.     

Free insulin growth factor-I and vascular endothelial growth factor in the vitreous fluid of patients with proliferative diabetic retinopathy

PURPOSE: To investigate the relationship between insulin-like growth factor-I (IGF-I) and vascular endothelial growth factor (VEGF) in the vitreous fluid of diabetic patients with proliferative diabetic retinopathy (PDR). DESIGN: Observational case-control study. METHODS: In a prospective study, 37 consecutive diabetic patients with PDR (14 type I and 23 type II diabetes mellitus) in whom a vitrectomy was performed were compared with 21 nondiabetic patients with other conditions requiring vitrectomy (control group). Free IGF-I and VEGF were measured by ELISA. RESULTS: Vitreal levels of both free IGF-1 and VEGF were higher in diabetic patients with PDR than in control subjects (P <.01, and P <.0001, respectively). After adjusting for total intravitreous protein concentration, VEGF (ng/mg of proteins) remained significantly higher in diabetic patients with PDR than in the control group (P <.0001), whereas free IGF-I (ng/mg of proteins) was lower in diabetic patients than in control subjects (P <.0001). The vitreous concentrations of VEGF were higher in patients with active PDR than in patients with quiescent PDR (P <.005), whereas vitreous free IGF-I was not related to PDR activity. Finally, we did not observe a correlation between the vitreal levels of free IGF-I and VEGF. CONCLUSIONS: We conclude that free IGF-I and VEGF are both increased, but not related, within the vitreous fluid of diabetic patients with PDR. In addition, our results support the current concept that VEGF is directly involved in the pathogenesis of PDR, whereas the precise role of free IGF-I remains to be established.     

Angiotensin II and vascular endothelial growth factor in the vitreous fluid of patients with proliferative diabetic retinopathy

AIMS: To investigate the correlation between the level of angiotensin II and vascular endothelial growth factor (VEGF) in the vitreous fluid and the severity of proliferative diabetic retinopathy (PDR). METHODS: During vitreoretinal surgery at the Tokyo Women's Medical University, vitreous fluid samples were obtained from 51 eyes of diabetic patients with PDR, six eyes of diabetic patients without retinopathy, and 16 eyes of non-diabetic patients with ocular disease (controls). The VEGF levels in vitreous fluid and plasma were determined by enzyme linked immunosorbent assay, while angiotensin II levels were measured by radioimmunoassay. RESULTS: The vitreous fluid levels of VEGF and angiotensin II were significantly higher in patients with PDR than in non-diabetic patients or diabetic patients without retinopathy (all p<0.0001). The vitreous fluid level of angiotensin II was significantly correlated with that of VEGF (p<0.0001), and the vitreous concentrations of both VEGF and angiotensin II were significantly higher in patients with active PDR than in those with quiescent PDR (p<0.0001 and p=0.0005, respectively). CONCLUSION: The authors found that both angiotensin II and VEGF levels were significantly higher in the vitreous fluid of patients with PDR than in that of non-diabetic patients or diabetic patients without retinopathy, and that the levels of both angiotensin II and VEGF were elevated in the active stage of PDR. These findings suggest that angiotensin II contributes to the development and progression of PDR in combination with VEGF.     

内皮抑素对视网膜新生血管中血管内皮细胞生长因子表达的影响

目的:观察内皮抑素(endostatin,ES)对视网膜新生血管中血管内皮细胞生长因子(vascular endothelial growth factor,VEGF)表达的影响。方法:通过缺氧的方法建立小鼠视网膜新生血管模型36只,随机分为高氧组及ES治疗组。ES治疗组小鼠在出氧箱后12,36h,玻璃体腔内注射ES1μL。另将生活在正常氧环境中的18只同龄小鼠作为正常对照。提取3组小鼠视网膜总RNA,通过RT-PCR方法定量检测VEGF在3组小鼠视网膜中的表达。结果:氧致视网膜病变视网膜中VEGF的表达升高,与正常对照组比较有统计学意义(高氧组与正常对照组比较P<0.05);ES作用后VEGF的表达减少,与给氧组比较有统计学意义(ES治疗组与高氧组比较P<0.05)。结论:ES作用机制可能是通过抑制VEGF mRNA基因的表达来抑制视网膜新生血管的形成。     

增生性糖尿病视网膜病变患者玻璃体内血管内皮生长因子的表达

目的 探讨增生性糖尿病视网膜病变(PDR)患者玻璃体内血管内皮生长因子(VEGF)的表达及相关影响因素.方法 病例对照研究.对50例(50只眼)接受玻璃体切除治疗的PDR患者进行分析.术中获取玻璃体标本,并用双抗体夹心酶联免疫吸附试验ELISA法对术中获取的玻璃体标本进行VEGF含量的定量检测,并分析增生性糖尿病视网膜病变患者玻璃体内VEGF的表达情况.平均随访9个月(6～26个月).用成组t检验比较PDR组和正常对照组玻璃体VEGF含量差异,以及硅油填充组与非硅油填充组VEGF含量差异.用方差分析方法比较PDR进展组、稳定组和好转组玻璃体VEGF含量有无差异.用单因素方差分析方法分析玻璃体VEGF含量对PDR术后疗效的影响.结果 PDR组玻璃体VEGF含量平均(592.4801±587.4267)ng/L,正常对照组玻璃体VEGF含量平均(131.3022±26.9192)ng/L.PDR组玻璃体VEGF含量高于对照组(t=3.2315,P＜0.05).术后PDR进展有10只眼(20%),稳定10只眼(20%),好转30只眼(60%).PDR进展组玻璃体VEGF含量显著高于PDR稳定和好转组(q=-3.3187,-4.0843;P＜0.05).术前未接受视网膜光凝组玻璃体VEGF含量显著高于接受全视网膜光凝或局部视网膜光凝组(q=-4.2187,-3.9672;P＜0.05).结论玻璃体VEGF表达与PDR严重程度有一定关系.术后PDR稳定或好转者玻璃体VEGF水平呈相对低表达.(中华眼科杂志,2009,45:206-209)     

Levels of vascular endothelial growth factor are elevated in the vitreous of patients with subretinal neovascularisation.

BACKGROUND: Vascular endothelial growth factor (VEGF) has been shown to play a major role in intraocular neovascularisation in ischaemic retinal diseases. Subretinal neovascularisation is an important cause of central visual loss, but little is known about the role of this growth factor in its pathogenesis. The aim of this study was to investigate the possible role of VEGF in the development of subretinal neovascularisation. METHODS: Undiluted vitreous samples were obtained from patients undergoing vitrectomy for removal of non-age-related subfoveal neovascular membranes (SFNM). For comparison vitreous from patients undergoing vitrectomy for idiopathic full thickness macular holes (FTMH) and proliferative diabetic retinopathy (PDR) was used. Indirect enzyme linked immunosorbent assay (ELISA), with an antibody directed against the conserved N-terminal region of human VEGF165, was used to determine vitreous levels of VEGF. The growth factor was also localised in the vitreous of patients with SFNM by western blot analysis. RESULTS: The mean (SE) VEGF concentration in the vitreous of patients with SFNM was 27.78 (2.22) ng/ml (n = 8), FTMH was 16.62 (0.9) ng/ml (n = 18), and PDR was 37.77 (3.28) ng/ml (n = 16). The differences between the PDR group and SFNM group versus the FTMH group were both significant (p = 0.0001 and p = 0.0015) as analysed by the Wilcoxon rank sum test). CONCLUSIONS: Vitreous levels of VEGF are significantly elevated in eyes with non-age-related subretinal neovascularisation compared with eyes with FTMH but not as elevated as in PDR. This suggests that VEGF is involved in subretinal angiogenesis.     

Correlation between angiotensin-converting enzyme, vascular endothelial growth factor, and matrix metalloproteinase-9 in the vitreous of eyes with diabetic retinopathy

PURPOSE: To investigate the involvement of angiotensin-converting enzyme (ACE) with angiogenic factors, vascular endothelial growth factor (VEGF), and matrix metalloproteinase (MMP)-9 in the vitreous of eyes with proliferative diabetic retinopathy (PDR). DESIGN: Observational case series. METHODS: Angiotensin-converting enzyme activity in the vitreous was measured by using a synthetic substrate for ACE. VEGF and MMP-9 concentrations were determined by enzyme-linked immunosorbent assay. RESULTS: Vitreous ACE activity was significantly higher in eyes with PDR (1.4 +/- 1.3 mU/ml) than in eyes with macular holes (0.22 +/- 0.11 mU/ml). Vitreous VEGF concentration in the eyes with PDR (1067 +/- 1076 pg/ml) was significantly higher than in eyes with macular holes (34 +/- 5.5 pg/ml). Vitreous MMP-9 concentration was also significantly higher in eyes with PDR (7.5 +/- 3.8 ng/ml) than in eyes with macular holes (4.2 +/- 1.4 ng/ml). Significant correlations between ACE and VEGF (P < .01) and between ACE and MMP-9 (P < .01) were observed in the vitreous of eyes with PDR. CONCLUSIONS: Angiotensin-converting enzyme was significantly correlated with angiogenic factors, VEGF and MMP-9, in the vitreous of eyes with PDR.     

Inhibition of vascular endothelial cell growth and trypsin activity by vitreous

Vitreous from bovine, human and chick embryo has been found to contain a trypsin inhibitory activity. Chymotrypsin-inhibitory activity was also identified in bovine and chick embryo vitreous. Following either ultrafiltration or Bio Gel P-10 chromatography, these activities appear in fractions having a molecular weight greater than 10000 MW (ultrafiltration) or greater than 13000 MW (P-10 void volume), and are separable from low molecular weight aortic endothelial cell growth inhibitory activity present either in the ultrafiltrate or P-10 retarded volume. Treatment of the trypsin inhibitory fraction with hyaluronidase had no effect on trypsin inhibition, nor did addition of hyaluronic acid inhibit trypsin. Chick embryo vitreous and hyalocyte-conditioned medium were found to contain aortic endothelial cell growth inhibitory activity in both the void volume and retarded volume fractions following Bio Gel P-10 chromatography. Both the 6200 MW bovine vitreous endothelial cell growth inhibitor and the high molecular weight chick embryo vitreous endothelial cell growth inhibitor (greater than 13000 MW) were similar, in that most of the activity did not bind to heparin linked to Sepharose CL-6B.