增生性玻璃体视网膜病变后的玻璃体后膜裂孔组织学改变

目的研究视网膜脱离伴有增生性玻璃体视网膜病变(PVR)的后膜裂孔组织学变化。方法3例孔源性视网膜脱离伴有PVR患者进行玻璃体手术同时获得后膜裂孔,应用免疫电镜和光镜技术对3个标本进行超微结构和细胞外基质蛋白、细胞中间丝蛋白标记观察。结果后膜裂孔膜组织有中等量细胞分布和丰富的胶原纤维。细胞主要有两种类型:含有色素颗粒的上皮样细胞和有明显微丝结构的梭形或肾形细胞,分散分布;Ⅰ和Ⅲ型胶原标记阳性。免疫组织化学光镜观察发现波形蛋白阳性标记最强,胶质原纤维酸性蛋白次之,细胞角质蛋白阳性标记较弱,而抗S-100、神经元特异性烯醇化酶以及巨噬细胞抗体标记偶有阳性标记细胞。结论研究提示视网膜脱离伴有PVR的后膜裂孔组织成分发生变化,除神经胶质细胞外,还有其他细胞参与,包括视网膜色素上皮细胞、成纤维样细胞,胶原类型也发生改变,后膜裂孔组织学上的变化与在玻璃体、视网膜表面形成的PVR膜一致。     

增生性玻璃体视网膜病变视网膜前膜超微结构及HGF受体表达

目的：研究增生性玻璃体视网膜病变视网膜前膜(epiretinal membrane,ERM)的超微结构及肝细胞生长因子(hepatocyte growth factor,HGF)受体的表达情况。方法：严重增生性玻璃体视网膜病变(PVR)患10例行玻璃体手术时取出视网膜前膜,透射电镜观察ERM的组成成分,免疫组化染色观察ERM组织中HGF。受体的表达。结果：ERM以上皮样细胞、成纤维样细胞为主并含有大量胶原成分。ERM标本中HGF受体集中在细胞分布密集区域表达,阳性细胞多是一些含色素颗粒的细胞。结论：视网膜色素上皮(RPE)细胞是PVR的主要参与细胞,HGF可能参与了PVR膜形成。     

增生性玻璃体视网膜病变发病机制的研究进展

增生性玻璃体视网膜病变(proliferative vitreoretinopathy，PVR)是在孔源性视网膜脱离或视网膜脱离复位术后，由于视网膜色素上皮细胞(RPE)及神经胶质细胞的增生和收缩，造成牵拉性视网膜脱离的病变。PVR的病理特征是细胞增生，RPE一旦开始增殖，就产生细胞生长因子，而这些细胞生长因子反过来又刺激细胞增殖。PVR的过程有多种细胞及因子的参与，参与PVR增生的细胞主要是RPE和视网膜神经胶质细胞；涉及PVR的生长因子有：PDGF、EGF、VEGF、TGF、FGF、IGF、HGF等。     

增生性玻璃体视网膜病变膜剥离标本的免疫组化研究

目的 探讨不同增生性玻璃体视网膜病变(PVR)的增殖特征。方法采用5种特异性抗体对12例PVR膜样本进行免疫组织化学研究。结果成纤维细胞、神经胶质细胞为参与PVR膜的主要细胞成分,视网膜色素上皮细胞(RPE)、巨噬细胞、纤维连接蛋白和新生血管也参与了PVR的病理过程。结论新生血管主要参与了增生性视网膜血管病变的病理过程。增殖膜中增殖的细胞、细胞外基质和血管成分参与了PVR的病理过程并起着不同的作用。     

增生性玻璃体视网膜病变中细胞因子的作用

细胞因子与PVR形成密切相关。HGF通过自分泌或旁分泌方式作用于RPE和神经胶质细胞，导致RPE活化和胶质细胞迁移；通过活化磷脂酰肌醇3激酶促使神经胶质细胞产生血管内皮生长因子（VEGF）。PDGF-AA通过与含有口亚基PDGFR作用促进PVR形成，但表达PDGFβ受体的细胞不能诱导PVR发生。肌样成纤维细胞聚集是PVR中导致牵拉性视网膜脱离的重要因素，其α平滑肌肌动蛋白的表达依赖于TGF-β2RⅡ及TGF-β1在增生膜中的表达。IL-6是PVR形成的危险因素，是视网膜脱离术后PVR复发的重要预测指标。TNF-α可与RPE表面TNF受体作用并通过MAPK通路诱导RPE细胞高表达α1与α5型整合素，进而促进RPE在PDGF介导下的细胞转移。MCP-1可能在PVR早期起重要作用。     

增生性玻璃体视网膜病变中细胞因子的作用

细胞因子与PVR形成密切相关。HGF通过自分泌或旁分泌方式作用于RPE和神经胶质细胞,导致RPE活化和胶质细胞迁移;通过活化磷脂酰肌醇3激酶促使神经胶质细胞产生血管内皮生长因子(VEGF)。PDGF-AA通过与含有α亚基PDGFR作用促进PVR形成,但表达PDGFβ受体的细胞不能诱导PVR发生。肌样成纤维细胞聚集是PVR中导致牵拉性视网膜脱离的重要因素,其α平滑肌肌动蛋白的表达依赖于TGF-β2RⅡ及TGF-β1在增生膜中的表达。IL-6是PVR形成的危险因素,是视网膜脱离术后PVR复发的重要预测指标。TNF-α可与RPE表面TNF受体作用并通过MAPK通路诱导RPE细胞高表达α1与α5型整合素,进而促进RPE在PDGF介导下的细胞转移。MCP-1可能在PVR早期起重要作用。     

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。     

用免疫荧光方法检测PVR增生膜中的EGFR

目的 观察表皮生长因子受体(EGFR)在增生性玻璃体视网膜病变(PVR)的增生膜(PRM)中的表达，并判断EGFR阳性细胞可能的来源。方法 PVR增生膜43例，按病程分为3期，采用免疫荧光法检测其EGFR表达并做定量分析；采用免疫荧光双重标记技术判定EGFR阳性细胞来源。结果 早期膜中阳性荧光总量约为中期膜的2．06倍，12例晚期膜均表达阴性；EGFR表达阳性的细胞，细胞角蛋白(cytokeratinpan)和巨噬细胞标记抗体(HAM-56)呈阳性表达。结论 视网膜色素上皮细胞及巨噬细胞具有EGFR，在PVR早期的发生发展起重要作用。     

增生性玻璃体视网膜病变增生膜中基质金属蛋白酶及其抑制剂的表达

目的研究增生性玻璃体视网膜病变（proliferative vitreoretinopathy，PVR）增生膜中基质金属蛋白酶（matrix metalloproteinases，MMP）及其抑制剂（tissue inhibitors of matrix metalloproteinases，TIMP）的表达。方法采用免疫组织化学技术的SP法，检测PVR增生膜和正常尸眼神经视网膜标本中MMP-2、MMP-9和TIMP-1的表达，光镜观察染色结果。结果41例PVR增生膜标本HE染色切片光镜下可见视网膜色素上皮（retinal pigment epithelial，RPE）细胞、神经胶质细胞、成纤维细胞、巨噬细胞等细胞成分，它们被大量细胞外基质所包绕。视网膜前膜中以RPE细胞为主要增生细胞，视网膜下膜中以神经胶质细胞为主要增生细胞。免疫组织化学染色结果显示：（1）25例PVR增生膜标本表达MMP-2，11例PVR增生膜标本表达MMP-9，14例PVR增生膜标本表达TIMP-1；（2）22例视网膜前膜标本表达MMP-2，3例视网膜下膜标本表达MMP-2，差异有显著性（P〈0．05）；（3）正常尸眼神经视网膜标本中未检测到MMP-2、MMP-9及TIMP-1的表达。结论细胞外基质与PVR增生膜的形成关系密切，MMP-2、MMP-9及TIMP-1在PVR形成过程中发挥重要作用，通过合理地调控MMP和TIMP的平衡，为预防和治疗PVR提供可能性理论依据。     

增生性玻璃体视网膜病变增生膜再塑型机制的研究

目的 观察不同病程增生性玻璃体视网膜病变（PVR）增生膜中不同细胞成分、细胞外基质（ECM）、基质金属蛋白酶（MMPs）及其抑制剂（TIMPs）随病程变化的规律，探讨PVR增生膜的再塑型机制。 方法 病程2个月至8年的孔源性视网膜脱离伴PVR患者的增生膜手术标本16例，用免疫组织化学方法标记增生膜中视网膜色素上皮（RPE）细胞、胶质细胞等不同的细胞成分，纤维连接蛋白（FN）、层粘连蛋白（l aminin）、Ⅰ～Ⅳ型胶原等不同ECM成分，以及MMPs（MMP2、MMP9）和TIMP1，分析不同病程PVR增生膜中各标记成分的变化以及与病程的相关性。 结果 随PVR病程延长，增生膜中RPE细胞、MMP2、FN表达逐渐减少(P=0.014，P=0.001，P=0.008)， 胶质细胞、Ⅰ、Ⅲ型胶原逐渐增多（P=0.022，P=0.001，P=0.008)，层粘连蛋白和Ⅱ、Ⅳ型胶原均有表达，但不随病程变化。RPE细胞、MMP2、纤维连接蛋白的表达与病程呈负相关，胶质细胞、Ⅰ、Ⅲ型胶原的表达与病程呈正相关；MMP2与FN变化呈正相关。MMP9、TIMP1始终都有表达，但不随病程变化。 结论 在PVR增生膜形成和发展的过程中，增生膜中的RPE细胞、胶质细胞、FN、Ⅰ、Ⅲ型胶原、MMP2参与了PVR的再塑型过程。 (中华眼底病杂志， 2006， 22： 308-312）     

Induction of proliferative vitreoretinopathy by a unique line of human retinal pigment epithelial cells

BACKGROUND: The most widely used models of proliferative vitreoretinopathy (PVR) rely on injection of cells into the vitreous of animals. Using retinal pigment epithelial (RPE) cells from human PVR membranes may produce a more accurate model of human PVR. We performed a study to determine whether human RPE cells derived from a single epiretinal membrane (ERM) are capable of inducing the same disease in the rabbit eye, and whether the induced ERMs had cellular components similar to those of human PVR membranes. METHODS: Cells were harvested from a human ERM obtained at surgery for PVR. RPE cells were cultured from the membrane and injected into the right eye of 24 New Zealand albino rabbits. The left eyes served as controls. The eyes were examined by indirect ophthalmoscopy over 4 weeks. The enucleated eyes were then examined by means of microscopy and histochemical analysis. RESULTS: By day 7, PVR had developed in all but 1 of the 24 experimental eyes, with 8 progressing to localized tractional retinal detachment. By day 21, localized tractional retinal detachment had developed in 17 eyes; 1 eye progressed to extensive tractional retinal detachment by day 28. Immunostaining showed that mostly RPE cells, but also myofibroblasts, glial cells and collagen, were present in the newly formed rabbit PVR membranes. INTERPRETATION: Human RPE cells cultured from a PVR membrane appear to be capable of inducing PVR in rabbits. The resultant ERMs are similar to those formed in human PVR and consist mainly of RPE cells.     

曲安奈德对PVR中增殖细胞抑制效果的观察

目的：探讨不同浓度曲安奈德对体外培养的视网膜前膜细胞、视网膜色素上皮细胞（RPE）及神经胶质细胞的抑制率差异,找出视网膜前膜细胞达最大抑制率时的最低有效药物浓度,用于指导临床外伤性增生性玻璃体视网膜病(PVR)防治。 方法：用四唑盐(MTT)比色法检测不同浓度曲安奈德(TA)对体外培养的视网膜前膜细胞、RPE及神经胶质细胞的作用。 结果：三种细胞对TA的反应不完全相同,其中视网膜前面细胞达最大抑制率时所需要TA的最低有效药物浓度0.8mg/mL,RPE细胞最大抑制率所需TA最低有效药物为0.4mg/mL,神经胶质细胞最大抑制率所需TA最低有效药物为0.6mg/mL;且抑制率与药物作用时间呈正相关性。 结论：TA能够有效抑制体外培养的视网膜前膜细胞、RPE细胞及神经胶质细胞的增生;抑制视网膜前膜细胞增殖所需TA药物浓度明显高于RPE和神经胶质细胞。     

The Immunological Studies on Proliferative Intraocular Disorders：Investigation of Activated Lymphocytes,Macrophages and HLA—DR

Purpose: To investigate whether activated B lymphocytes ( CD23 ) , activated T lymphocytes (CD25) ,macrophages(CD68) and human leucocyte antigen class II antigen (HLA- DR) were existed in epiretinal membranes (ERMs) and subretinal membranes (SRMs) of prolif-erative intraocular disorders (PID).Methods : Twenty specimens of ERMs from rhegmatogenous retinal detachment with prolifera-tive vitreoretinopathy( PVR), traumatic PVR and secondary traction retinal detachment, and two specimens of SRMs from rhegmatogenous retinal detachment with PVR and traumatic PRV were studied using immunohistochemical staining.Results: CD68 and HLA - DR were found in all specimens, CD23 and CD25 in 4 cases of ERMs and in 1 case of SRMs, respectively.Conclusions : 1. The ERMs and SRMs of different etiology shared a common basis of inflammation and immunopathology. 2. There would be secondary cellular and humoral immunity in the ERMs and the SRMs of PID. Eye Science 1998; 14:35 - 40.     

肝细胞生长因子受体在视网膜前膜及培养的人视网膜色素上皮细胞中的表达

目的 观察增生性玻璃体视网膜病变(proliferative vitreoretinopathy,PVR)C、D两级视网膜前膜 (epiretinal membranes, ERM)和培养的人视网膜色素上皮(retinal pigment epithelium, RPE)细胞中肝细胞生长因子受体（hepatocyte growth factor receptor, HGFR）的表达情况。 方法 采用免疫组织化学染色方法对15例复杂孔源性视网膜脱离患者玻璃体切割术中剥离的ERM以及培养的人RPE细胞中HGFR的表达情况进行观察。 结果 在6例PVR C级和9例PVR D级ERM标本中分别有5、7例呈HGFR阳性表达；培养的RPE细胞胞浆中HGFR呈阳性表达。 结论 肝细胞生长因子有可能参与了 PVR的病理过程。 (中华眼底病杂志, 2002, 18: 221-223)     

Upregulation of RAGE and its ligands in proliferative retinal disease

We sought to study the presence of the receptor for advanced glycation endproducts (RAGE) and its ligands, advanced glycation endproducts (AGEs), S100/calgranulins and amphoterin (high mobility group box 1 protein; HMGB1), in the vitreous cavity and epiretinal membranes (ERMs) of eyes of patients with proliferative diabetic retinopathy (PDR) and proliferative vitreoretinopathy (PVR). Undiluted vitreous specimens were collected from 30 eyes of 30 patients undergoing pars plana vitrectomy for repair of retinal detachment (RD) secondary to PDR (n = 15) or PVR (n = 15). The vitreous samples obtained from 10 eyes undergoing macular hole repair were used as controls. Epiretinal membranes were obtained from eight eyes with PDR and from 10 eyes with PVR. The levels of AGEs in the vitreous were measured using ELISA. The vitreous levels of soluble RAGE (sRAGE), S100/calgranulins and amphoterin were measured using Western blot analyses. The localization of RAGE and its ligands in ERMs was determined with immunohistochemistry. The vitreous levels of sRAGE were significantly increased in both PDR and PVR (p < or = 0.05) compared to control vitreous. In both PDR and PVR, the vitreous levels of AGEs (p < or = 0.01), S100/calgranulins (p < or = 0.05), and amphoterin (p < or = 0.01) were also elevated compared to control eyes. Expression of RAGE was detected in six of eight ERMs from eyes with PDR and eight of 10 ERMs from eyes with PVR. Many cells expressing RAGE also expressed vimentin, suggesting a glial cell origin. Ligands for RAGE were also detected in ERMs, with AGEs detected in five eyes with PDR and eight eyes with PVR. Similarly, S100 and amphoterin ERM expression was observed in six eyes with PDR; these ligands were also expressed in ERMs from eyes with PVR (8 and 7 cases, respectively). We conclude that RAGE and its ligands are increased in the vitreous cavity of eyes with PDR and PVR and are present in ERMs of eyes with these proliferative retinal disorders. These findings suggest a role for the proinflammatory RAGE axis in the pathogenesis of proliferative retinal diseases.     

孔源性视网膜脱离患者视网膜表面膜组织的体外培养

目的 建立体外培养增生性玻璃体视网膜病变（PVR）患者病变膜组织的方法，为提供PVR研究奠定基础。方法 孔源性视网膜脱离伴有PVR分级D级患者进行玻璃体切割术同时获取其视网膜表面膜病变（ERM）标本，患者病程7～120天；ERM膜片应用切块贴皿法进行接种，培养皿短时间斜置使ERM组织贴壁，每3～4天换培养液1次，观察记录膜组织中细胞的迁出、分裂和增生状况，部分病变标本进行免疫细胞化学鉴定。结果 7例患者的ERM组织提供体外原代培养，培养期间膜片色素变淡，其中3例患者的膜组织有细胞从膜片边缘向外迁移，生长缓慢，迁出的细胞呈现两种形态：铺路石样和梭形。cytokeratin染色呈阳性。结论 患者ERM体外培养方法可行，细胞生长情况与多因素有关。ERM中的细胞主要是RPE细胞，其增殖能力有可能评估手术预后。     

微小RNA调控增生性玻璃体视网膜病变的研究进展

增生性玻璃体视网膜病变(PVR)是一个眼部组织的创伤修复和纤维化过程,其特征性改变是在玻璃体腔和(或)视网膜表面形成由细胞外基质(ECM)和各种类型的细胞组成的视网膜前膜(ERM),ERM收缩形成视网膜皱褶,并牵拉视网膜引起视网膜脱离(RD)。视网膜色素上皮(RPE)细胞发生上皮-间质转化(EMT)和ECM累积是ERM形成的重要病理机制。转化生长因子-β(TGF-β)等诱导RPE细胞发生EMT,细胞失去上皮表型、细胞间黏附减弱和表达间充质表型。由间充质细胞分化而成的成纤维细胞样细胞分泌ECM等成分,与神经胶质细胞、成纤维细胞等共同形成ERM。RPE细胞的EMT过程受许多细胞因子/生长因子、转录因子及微小RNA(microRNA,miRNA)等的调控,研究证明miRNA是一类新型且功能强大的调节基因,在PVR的EMT过程中起着重要调控作用。本文将近年来miRNA调控PVR的作用机制和干预性治疗研究进行综述。     

Retina-specific expression of PDGF-B versus PDGF-A: vascular versus nonvascular proliferative retinopathy

PURPOSE: Platelet-derived growth factor (PDGF) has been implicated in vascular proliferative retinopathies, such as diabetic retinopathy, and in nonvascular retinopathies, such as proliferative vitreoretinopathy. Traction retinal detachment is a central feature of both types of disease. Hemizygous rhodopsin promoter/PDGF-B (rho/PDGF-B) transgenic mice exhibit proliferation of vascular cells, glia, and retinal pigmented epithelial (RPE) cells, resulting in traction retinal detachment. Hemizygous rho/PDGF-A transgenic mice show mild proliferation of glial cells and no traction retinal detachments. This study was undertaken to determine whether higher levels of endogenously produced PDGF-A in the retinas of mice result in retinal detachment. METHODS: To achieve high-level expression of PDGF-A in the retina, homozygous rho/PDGF-A (rho/PDGF-AA) mice were generated. The phenotype of these mice was compared with that of homozygous rho/PDGF-B (rho/PDGF-BB) mice and double hemizygous rho/PDGF-B-rho/PDGF-A (rho/PDGF-AB) mice. RESULTS: Rho/PDGF-BB and rho/PDGF-AB mice showed a phenotype similar to that previously described in rho/PDGF-B mice. There was extensive proliferation of glial and vascular cells, resulting in fibrovascular membranes that detached the retina. PDGF-AA mice showed extensive proliferation of glial cells and traction retinal detachment. CONCLUSIONS: High retinal expression of PDGF-A results in extensive proliferation of glial cells and traction retinal detachment without vascular cell involvement, similar to proliferative vitreoretinopathy in humans. High retinal expression of PDGF-B results in traction retinal detachment from proliferation of both vascular and nonvascular cells, similar to diabetic retinopathy in humans.     

TGF-beta1, TGF-beta receptor II and ED-A fibronectin expression in myofibroblast of vitreoretinopathy

PURPOSE: Formation of scarlike epiretinal membranes (ERMs) constitutes potentially the end stage of evolution of proliferative vitreoretinopathy (PVR) and proliferative diabetic retinopathy (PDR). Among various cellular populations, ERMs contain cells with contractile features typical of myofibroblasts. The current study was conducted to investigate the presence of transforming growth factor (TGF)-beta1, TGF-beta receptor II (RII) and ED-A fibronectin (FN), the main inducers of myofibroblastic differentiation in ERMs in PDR and PVR. METHODS: Samples of ERM were obtained from 23 patients during microsurgery for PVR or PDR. Electron microscopy, immunohistochemistry, and confocal microscopy with antibodies recognizing beta-smooth muscle (SM) actin, desmin, TGF-beta1, TGF-beta receptors I and II, and ED-A FN were performed. RESULTS: alpha-SM actin was detected in all ERMs, whereas desmin was present in 50% of the cases. ED-A FN was expressed in all ERMs in close relation with alpha-SM actin-positive myofibroblasts. In addition, TGF-beta1 and TGF-beta R II were always present, TGF-beta RII being expressed in both alpha-SM actin-positive and negative fibroblastic cells. CONCLUSIONS: Myofibroblast accumulation is a key event in ERM-associated traction retinal detachment occurring during PVR and PDR. The current results suggest that the presence of alpha-SM actin-positive myofibroblasts is probably dependent on the concomitant neoexpression of TGF-beta1, TGF-beta RII, and ED-A FN. The results furnish new data on the mechanism of alpha-SM actin stimulation in fibroblasts in a human pathologic setting.