Platelet derived growth factor and fibroblast growth factor basic levels in the vitreous of patients with vitreoretinal disorders

AIM—To determine the potential role of basic fibroblast growth factor (bFGF) and platelet derived growth factor (PDGF) in the pathogenesis of proliferative vitreoretinopathy (PVR).
METHODS—An enzyme linked immunosorbent assay technique was used to determine the quantities of bFGF and PDGF in 38 vitreous samples taken from patients undergoing trans pars plana vitrectomy (PPV) for a variety of vitreoretinal disorders.
RESULTS—bFGF levels ranged from undetectable to 318.7 pg/ml. The mean concentration was 27.57 pg/ml. PDGF levels ranged from undetectable to 160 pg/ml, the mean concentration being 40.06 pg/ml. Eight of 13 patients with clinical evidence of retinal detachment (RD) and PVR had significantly elevated bFGF concentrations, whereas only one of 11 patients with RD and no PVR had detectable bFGF; seven of eight patients with RD and PVR had elevated PDGF concentrations in the vitreous, whereas all 10 patients with RD and no PVR had no detectable vitreous PDGF. Eight patients with vitreous haemorrhage had raised PDGF concentrations, and the levels were particularly high (>120 pg/ml) in those two patients with active neovascularisation. Two out of nine patients with vitreous haemorrhage had raised bFGF levels.
CONCLUSIONS—bFGF and PDGF concentrations are elevated in PVR even in the absence of vitreous haemorrhage, and not in patients with RD uncomplicated by PVR. This observation suggests that both cytokines may be involved in the pathogenesis of PVR.

Keywords: basic fibroblast growth factor; platelet derived growth factor; proliferative vitreoretinopathy     

结缔组织生长因子mRNA在增生性糖尿病视网膜病变视网膜前纤维血管膜中的表达

目的：观察结缔组织生长因子（connective tissue growth factor ,CTGF)mRNA在增生性糖尿病视网膜病变（proliferative diabetic retinopathy,PDR)纤维血管性视网膜前膜中的表达。方法：采用原位杂交方法，对玻璃体切割术获得的6例PDR的纤维血管性膜进行CTGF mRNA的检测。结果：2例纤维血管性膜标记为阳性染色（＋＋），4例为强阳性染色（＋＋＋），每例标本随机计数100个细胞中的阳性细胞数，取6例标本的平均值，计算平均阳性率为77％，阳性细胞多见于成纤维细胞样细胞和新生血管的血管内皮细胞。结论：PDR纤维血管性膜形成过程中成纤维样细胞及血管内皮细胞在TGF－β等生长因子的刺激下，CTGF mRNA显著上调，CTGF参与了PDR纤维血管性膜的形成和发展。     

PDR、PVR增生膜组织超微结构及生长因子受体表达生物学特点的比较研究

目的比较研究增生性玻璃体视网膜疾病PVR及PDR增生膜组织超微结构与生长因子受体表达的生物学特点。方法PVR、PDR增生膜组织标本29例,通过电镜与免疫组织化学方法比较研究PVR与PDR增生膜组织超微结构、细胞膜上TGFβR1和EGF受体表达的生物学特点。结果(1)PVR、PDR增生膜中细胞类型主要是RPE细胞、纤维细胞、纤维母细胞样细胞和巨噬细胞;PVR与PDR膜组织超微结构不同。(2)TGFβR1受体在PDR增生膜组织中的表达,其平均光强度(P<0.01)与平均阳性面积(P<0.05)均强于PVR增生膜组织,EGF受体在PDR增生膜组织中(13/14,92.8%)表达较PVR增生膜组织中(3/15,20%)明显(P<0.05)。(3)无论PDR、PVR增生膜组织TGFBRI受体在病程>6个月的增生膜组织中的表达,明显强于小于6个月的标本(P=0.002),而EGF受体在病程2-6个月的膜组织中表达平均面积(P=0.024)明显小于其他病程的膜组织。结论PDR、PVR增生膜组织,其超微结构与生长因子表达的生物学特点不同。     

Expression of autotaxin and acylglycerol kinase in proliferative vitreoretinal epiretinal membranes

Purpose: Lysophosphatidic acid (LPA)/LPA₁ receptor pathway is involved in inflammation, angiogenesis and fibrosis. This study was conducted to analyse the expression of LPA‐producing enzymes, autotaxin (ATX) and acylglycerol kinase (AGK) and LPA₁ receptor, in proliferative diabetic retinopathy (PDR) and proliferative vitreoretinopathy (PVR) epiretinal membranes. Methods: Nine active and 13 inactive membranes from patients with PDR and 21 membranes from patients with PVR were studied by immunohistochemistry. Results: In PDR membranes, vascular endothelial cells expressed ATX and AGK in 16 and 19 membranes, respectively. Stromal cells expressed ATX and AGK in 19 and 22 membranes, respectively. Immunoreactivity for LPA₁ receptor was noted in vascular endothelial cells and stromal cells in the five membranes stained for LPA₁ receptor. Numbers of blood vessels and stromal cells expressing CD34, ATX and AGK were significantly higher in active membranes than in inactive membranes. Significant correlations were detected between number of blood vessels expressing the panendothelial cell marker CD34 and number of blood vessels and stromal cells expressing ATX and AGK. In PVR membranes, spindle‐shaped myofibroblasts expressing α‐smooth muscle actin co‐expressed ATX, AGK and LPA₁ receptor. Conclusions: The LPA/LPA₁ receptor pathway may be involved in inflammatory, angiogenic and fibrotic responses in proliferative vitreoretinal disorders.     

Changes in vitreous VEGF, bFGF and fibrosis in proliferative diabetic retinopathy after intravitreal bevacizumab

AIM: To evaluate the relationship between intravitreal bevacizumab (IVB) treatment and the levels of vitreous vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF) and vitreous-retina surface fibrosis in patients with proliferative diabetic retinopathy (PDR). METHODS: This study was a prospective, open-label, controlled, randomized clinical trial. Sixty-eight eyes of PDR patients (n=53) and macular hole patients (n=15) were enrolled in this study. Thirty-four eyes of the PDR patients received IVB before vitrectomy. Twenty-three of the 34 PDR patients received IVB treatment 5d before vitrectomy (subgroup a), and 11 of the 34 PDR patients received IVB treatment greater than 2wk prior to vitrectomy (subgroup b). Nineteen of the PDR patients did not receive IVB treatment at any time prior to vitrectomy. The levels of bFGF and VEGF in vitreous samples were measured using enzyme-linked immunosorbent assay (ELISA) and the degree of vitreoretinal fibrosis was characterized using clinical data and data obtained intra-operatively. RESULTS: In PDR patients, VEGF and bFGF levels were significantly increased compared to non-PDR (control) subject’s eyes (P<0.01). In PDR patients, vitreous VEGF levels were significantly decreased following IVB treatment compared to PDR patients that did not receive IVB treatment (P<0.01). The degree of vitreoretinal fibrosis was significantly increased in subgroup b compared to subgroup a(P<0.05) and to patients that did not receive IVB (P<0.05). Vitreous bFGF levels were significantly greater in subgroup b than subgroup a (P<0.01) or in patients who did not receive IVB treatment (P<0.05). A Spearman’s rank correlation test indicated that higher levels of vitreous bFGF, but not VEGF, correlated with the degree of vitreoretinal fibrosis. CONCLUSION: We found that bFGF levels increase in PDR patient’s vitreous after IVB treatment longer than two weeks prior to vitrectomy and correlated with the degree of fibrosis after IVB treatment. These findings suggest vitreous fibrosis is increased in PDR patients after IVB treatment may be due to increased levels of bFGF.     

表皮生长因子受体在增生性玻璃体视网膜病变视网膜周膜中的表达

目的 观察表皮生长因子受体(epidermalgrowthfactor receptor，EGFR)在增生性玻璃体视网膜病变(proliferative vitreoretinopathy，PVR)增生膜中的表达及其与病程的关系。方法 玻璃体手术中取出的PVR增生膜43例，按病程分为早期膜(＜2个月)，中期膜(2—6个月)和晚期膜(＞6个月)，用免疫组织化学及原位杂交技术从蛋白质及mR—NA水平检测EGFR的表达。结果 EGFR蛋白分子在早期膜中呈强阳性表达，中期膜中呈弱表达，晚期膜中呈阴性。EGFR基因仅在早期膜中呈阳性表达。结论 EGFR主要存在于PVR的早期膜中，可能介导有丝分裂信号对PVR的发生发展所起的重要调控作用。     

Vitronectin and proliferative intraocular disorders. II. Expression of cell surface receptors for fibronectin and vitronectin in periretinal membranes

Several cell types participate in the formation of vitreoretinal traction membranes in proliferative intraocular disorders. The communication between these cells involves hormones, growth factors, and the interaction with extracellular matrix molecules. We have previously demonstrated a partial colocalisation of two potent mediators of cell attachment, fibronectin and vitronectin, in periretinal membranes from patients with proliferative vitreoretinopathy (PVR). We found a similar pattern of vitronectin and fibronectin deposition in proliferative diabetic retinopathy (PDR) (n = 6). Now we show the expression of the corresponding cell surface receptors, integrins, for fibronectin and vitronectin by proliferating cells in 22 periretinal membranes, including traumatic (n = 8) and idiopathic (n = 8) PVR as well as PDR membranes (n = 6). Integrins are membrane receptors for extracellular matrix macromolecules which are involved in such basic biological phenomena as embryogenesis and metastasis. Future studies on the pathogenesis of vitreoretinal proliferation will have to focus on the initiation, maintenance, and regulation of this intercellular communication network involving attachment proteins and integrins.     

新鲜和冷冻保存的人羊膜中bFGFmRNA的检测

目的 研究新鲜和冷冻保存羊膜中是否存在碱性成纤维细胞生长因子(bFGF)mRNA。方法 从新鲜和保存1周羊膜中用常规的酚、氯仿法抽提总RNA，RT-PCR一步法扩增。PCR产物回收后与PBS质粒同时以EcoRⅠ、HindⅢ双酶切．将酶切产物通过T4DNA连接酶连接，重组质粒。将重组质粒导入感受态的DH5a大肠杆菌中．筛选阳性菌落抽提质粒后行酶切鉴定．选取阳性克隆测序。结果 RT-PCR产物电泳．可见单一清晰的条带，约410bp。重组质粒酶切后见二条带，小带约410bp，与插入片段一致，测序结果表明目的片段与bFGF的序列完全相同。结论 新鲜和冷冻保存的羊膜组织中存在bFGF的mRNA。     

血清中VEGF和bFGF水平与糖尿病视网膜病变关系的临床研究

目的:探讨糖尿病视网膜病变患者血清中血管内皮细胞生长因子(VEGF)、碱性成纤维细胞生长因子(bFGF)水平与糖尿病视网膜病变的关系。方法:收集患有糖尿病视网膜病变白内障患者的血清样本,并以无糖尿病白内障患者的血清样本作为对照,用ELISA方法测定血清中bFGF,VEGF的含量。结果:对照组12例血清样本bFGF含量为(34.95±9.62)ng/L,VEGF含量为(150±20)μg/L;单纯型DR组中10例bFGF含量为(52.6±7.54)ng/L,VEGF含量为(330±50)μg/L;增生型DR组7例bFGF含量为(64.929±11.917)ng/L,VEGF含量为(580±50)μg/L。和对照组相比糖尿病患者血清中bFGF,VEGF的含量均显著增加(P<0.01),并且随着DR病情的进展而增高(P<0.05);血清中VEGF与bFGF之间呈正相关(r=0.419,P<0.01)。结论:VEGF,bFGF参与了DR的发生发展,并与DR后期新生血管形成关系密切。     

PDR中血管内皮生长因子与纤维化相关细胞因子的相关性

血管内皮生长因子在视网膜新生血管生成中是必不可少的重要诱导因子。增殖性糖尿病视网膜病变（ proliferative diabetic retinopathy, PDR）患者视网膜新生血管形成后,随着病情进一步加重,可造成纤维血管膜形成、视网膜前膜的纤维化加重,造成牵拉性视网膜脱离。目前研究认为,细胞因子具有促进成纤维细胞增殖、移动、黏附和分泌细胞外基质的功能,在糖尿病环境状态下,转变至促纤维化状态,导致了细胞外基质的积聚和纤维化。本文对血管内皮生长因子与纤维化相关细胞因子相关性的研究现状予以综述。     

肝细胞生长因子受体在增生性糖尿病视网膜病变视网膜前膜中的表达

目的 肝细胞生长因子(hepatocyte growth factor receptor,HGF)是具有多种生物活性的细胞因子,通遇与受体结合转导生物学效应,发挥促细胞增生、促运动和促形态发生等作用。本实验目的在于观察增生性糖尿病视网膜病变(proliferative diabetic retinopathy,PDR)不同时期视网膜前膜(epiretinal membranes,ERM)中HGF受体的表达情况。方法 13例PDR患者行玻璃体切除术剥离视网膜前膜。采用免疫组化染色方法观察HGFR在ERM中的表达情况。结果 在7例Ⅳ、Ⅴ期ERM标本中,5例HGF受体阳性表达,阳性表达率为71.4％;6例PDRⅥ期增殖前膜标本中,有5例HGF受体阳性表达,阳性表达率为83.3％。HGF受体阳性表达率与PDR分期无显著性差别(P>0.05)。结论 HGF受体在PDR视网膜前增生膜中表达,提示HGF可能参与了PDR增生膜的形成。     

miRNA-15b和成纤维细胞生长因子2(FGF2)在增生型糖尿病视网膜病变患者玻璃体及视网膜增生膜组织中的表达

目的探讨增生型糖尿病视网膜病变(PDR)患者玻璃体、视网膜增生膜组织中miRNA-15b(miR-15b)和成纤维细胞生长因子2(FGF2)的表达情况,并进行相关性研究,初步探讨PDR发生原因。方法选取2016年9月至2019年10月期间在重庆市开州区人民医院眼科住院的PDR患者80例(80眼)作为PDR组,收集其玻璃体及视网膜增生膜组织;选取同期因眼外伤在本院眼科行眼球摘除术的2型糖尿病患者21例(21眼)作为对照组,收集其玻璃体及视网膜组织;通过实时荧光定量聚合酶链式反应(qRT-PCR)法检测玻璃体及视网膜组织/视网膜增生膜组织中miR-15b水平,分别通过酶联免疫吸附法(ELISA)、Western blot法检测玻璃体、视网膜组织/视网膜增生膜组织中FGF2蛋白水平表达;通过Pearson相关性分析法分析对照组玻璃体及视网膜组织以及PDR组患者玻璃体及视网膜增生膜组织中miR-15b与FGF2水平相关性;通过二元Logistic回归分析影响糖尿病患者发生PDR的危险因素。结果 PDR组糖尿病病程显著长于对照组(P<0.001)。PDR组玻璃体及视网膜增生膜组织中miR-...     

成纤维细胞生长因子21在糖尿病视网膜病变中的研究进展

成纤维细胞生长因子21(FGF21)作为新近发现的内源性调节因子,是研究的热点,其在糖尿病视网膜病变中起着重要的作用,因此近年来FGF21逐渐受到人们关注。本文对FGF21的分子结构、生物学功能及其与炎症反应的关系、在糖尿病视网膜病变病理进程中的作用等方面内容进行探讨。     

无环鸟苷滴眼液联合碱性成纤维细胞生长因子滴眼液治疗单纯疱疹病毒性角膜炎疗效观察

目的 评价无环鸟苷滴眼液联合碱性成纤维细胞生长因子（重组bFGF）滴眼液治疗单纯疱疹病毒性角膜炎的疗效。方法 回顾性分析单纯疱疹病毒性角膜炎96例（96只眼）,以抗病毒药物方法治疗为对照组,共46例;抗病毒药物联合碱性成纤维细胞生长因子滴眼液为治疗组,共50例。比较两组疗效。结果两组痊愈率比较,有显著性差异（P〈0.05）。结论 无环鸟苷滴眼液联合碱性成纤维细胞生长因子滴眼液组疗效优于对照组。     

VEGF在PDR中的表达及作用的研究

目的:研究增殖性糖尿病视网膜病变(proliferative diabetic retinopathy,PDR)患者血液、房水、玻璃体中血管内皮生长因子(vascular endothelial growth factor,VEGF)含量的变化,探讨VEGF与PDR的关系,为抗VEGF药物治疗的给药途径及剂量等提供理论依据。方法:采用双抗体夹心酶联免疫吸附测定法定量检测无糖尿病视网膜病变(NDR)组,单纯性糖尿病视网膜病变(BDR)组,增殖性糖尿病视网膜病变(PDR)组患者和正常对照组血浆中VEGF含量,还检测PDR患者房水、玻璃体中和正常对照组房水、玻璃体中VEGF含量,并进行综合分析。试剂盒购自美国R&D公司,其质量和灵敏度相对较高。结果:PDR组房水中VEGF含量有增高趋势,但与正常对照组比较,无统计学差异(P>0.05)。PDR患者玻璃体中VEGF含量明显增高,与正常对照组比较差异非常显著(P<0.01)。PDR组自身血浆、房水、玻璃体中VEGF含量比较有逐渐增高趋势,三者之间有显著性差异(P<0.01)。正常对照组血浆、房水、玻璃体中VEGF含量三者之间无显著性差异(P>0.05)。血浆VEGF含量在正常对照组中最高,而玻璃体中VEGF含量在PDR患者中最高。结论:PDR患者眼内尤其是玻璃体中VEGF含量大幅度增高,可能对促进DR发展恶化起了关键性的作用。在正常人,VEGF更多地存在于血浆中发挥其生物学效应。在严重DR患者中,玻璃体中异常地出现大量VEGF,推测来自缺血缺氧的视网膜,并可能有向眼前段扩散的趋势。     

结缔组织生长因子在增生性玻璃体视网膜病变增生膜中的表达

目的观察结缔组织生长因子(CTGF)在增生性玻璃体视网膜病变(PVR)增生膜中的表达,并鉴别增生膜中阳性细胞来源。方法采用SABC免疫组织化学方法对玻璃体切割手术获得的43例PVR增生膜进行CTGF蛋白的检测,并采用免疫荧光双标记技术在阳性表达的增生膜和正常人眼球标本中判定CTGF阳性细胞来源。结果免疫组织化学显示PVR增生膜中阳性细胞形态多是一类胞体为长圆形或多角形,胞浆丰富的上皮样细胞。17例C2-C3级膜中12例阳性,26例D1-D3级膜中19例阳性,其染色反应为阴性"-"、弱阳性" "、阳性"2 "和强阳性"3 "的分别有5、3、6、3例和7、5、8、6例,总阳性率分别为70.6%和73.1%。统计学分析CTGF阳性表达与膜分级问无相关性(P>0.1)。免疫荧光双标记法显示PVR增生膜中有RPE、巨噬细胞、神经胶质细胞,CTGF阳性细胞来源于RPE细胞;正常眼球RPE层不表达CTGF。结论正常眼球RPE层不表达CTGF,PVR形成过程中RPE细胞在TGF-β1等生长因子的刺激下,CTGF表达上调,表明CTGF参与了PVR增生膜的形成和发展。     

Recent developments in our understanding of how platelet-derived growth factor (PDGF) and its receptors contribute to proliferative vitreoretinopathy

Proliferative vitreoretinopathy, a disease process occurring in the setting of a rhegmatogenous retinal detachment, is thought to develop as a result of exposure of retinal cells to vitreous. Vitreous contains many growth factors, and platelet-derived growth factor (PDGF) has been considered a major contributor to PVR. Evaluation of both PDGF and PDGF receptors (PDGFRs) as potential therapeutic targets in the context of a rabbit model of PVR revealed that PDGFR-based approaches protected from PVR, whereas neutralizing PDGFs was a much less effective strategy. The basis for these observations appears to reflect that fact that the PDGFR could be activated by a wide spectrum of vitreal agents that are outside of the PDGF family. Furthermore, blocking signaling events by which the non-PDGFs indirectly activated PDGF α receptor (PDGFRα) protected rabbits from developing PVR. These studies demonstrate that the best therapeutic targets for PVR are not PDGFs, but PDGFRα and certain signaling events required for indirectly activating PDGFRα.     

抗VEGF辅助治疗增殖性糖尿病视网膜病变对眼内炎症因子水平的影响

目的:探讨玻璃体切除(PPV)术前辅助应用抗血管内皮生长因子(VEGF)药物(康柏西普)对增殖性糖尿病视网膜病变(PDR)患者玻璃体液中炎症细胞因子水平的影响。方法:收集2017-06/2018-01于南京医科大学第一附属医院眼科就诊并确诊的PDR患者49例49眼,随机分为两组,no-IVC组患者25例25眼PPV术前未接受玻璃体腔注射治疗,IVC组患者24例24眼PPV术前5～7d接受玻璃体腔注射康柏西普治疗。纳入患者均于PPV术前收集玻璃体液样本,并采用Luminex液相悬浮芯片技术检测血管内皮生长因子A(VEGF-A)、单核细胞趋化因子(MCP-1)及炎症细胞因子的水平。结果:与no-IVC组相比,IVC组患者玻璃体液中VEGF-A水平显著降低(P<0.001),炎症细胞因子IL-6(P=0.004)、IL-8(P=0.002)、IL-18(P=0.04)、TNF-α(P=0.03)水平显著升高,其余炎症细胞因子水平均无显著差异。结论:PPV术前辅助玻璃体腔注射康柏西普可有效降低PDR患者玻璃体液中VEGF-A水平,但对炎症细胞因子水平的影响有限。     

糖尿病性视网膜病变的手术时机及疗效的探讨

目的　探讨增生性糖尿病性视网膜病变(PDR)的手术时机与疗效的关系。方法　我院1998年8月～2 0 0 3年8月间行玻璃体视网膜手术的增生性糖尿病视网膜病变44例(5 6眼) ,分为增生早期组(糖尿病视网膜病变合并玻璃体积血或既往曾作全视网膜光凝治疗) 2 4眼,增生晚期组(广泛血管纤维膜增生合并牵引性视网膜脱离) 3 2眼,进行回顾性分析。结果　经过3月～3年随访(平均11.3月) ,增生早期组2 4眼均有不同程度的视力改善,0 .3者2 0眼(83 .3 3 % ) ,最佳视力达1.0 ;增生晚期组15眼视力有所提高,最佳视力为0 .4,4眼视力下降(其中两眼继发新生血管性青光眼而黑目蒙)。结论　正确把握糖尿病PDR玻璃体切除术手术时机,同时玻璃体切除术中行足量有效的眼内光凝,才能提高疗效降低糖尿病眼病的致盲率。     

散血明目片对兔tPVR玻璃体中生长因子和细胞间粘附分子浓度的影响

目的探讨活血利水之散血明目片对外伤性增生性玻璃体视网膜病变（traumatic proliferative vitmoretinopathy，tPVR）玻璃体中碱性成纤维细胞生长因子（basic fibroblasts growth factors，bFGF）、表皮生长因子（epidermal growth factor，EGF）和细胞间粘附分子-1（intercellular adhesion molecule-1，ICAM—1）浓度的影响及防治PVR的作用机理。方法50只家免随机抽取40只，造成外伤性PVR模型．随机分成散血明目片组、桃红四物汤组、猪苓散组、模型组，另10只为空白组。连续灌胃1月后，检测玻璃体中bFGF、EGF和ICAM-1的含量，检眼镜眼底检查并取眼球壁标本作组织病理学观察。结果散血明目片组玻璃体中bFGF、EGF、ICAM—1含量低于模型组，2者有显著性差异（P〈0．01）．散血明目片组EGF含量与空白组比较差异无显著性（P〉0．05），bFGF含量高于空白组（P〈0．05），桃红四物汤组与猪苓散组bFGF、EGF、ICAM-1浓度均低于模型组，且均高于空白组，有显著性差异（P〈0．01），桃红四物汤组与猪苓散组bFGF、EGF、ICAM-1浓度高于散血明目片组，有显著差异（P〈0．01）。各治疗组ICAM-1含量高于空白组，有显著性差异（P〈0、01）。结论活血利水之散血明日片有可能通过降低玻璃体中bFGF、EGF和ICAM-1的浓度，抑制增殖细胞的过度增生从而防治PVR形成和发展。