活血利水法对兔外伤性PVR增殖膜上EGFmRNA表达的影响

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 proliferrative vitreoretinopathy,tPVR)增殖膜(epiretinal membrane,ERM)中EGFmRNA表达的影响及防治PVR的作用机制。方法:将40只成年有色家兔随机抽取32只,制作tPVR模型,随机分成模型组(B组)、活血利水组(C组)、活血化瘀(D组)、利水明目组(E组),另8只为空白组(A组)。连续灌胃30d后,观察眼底PVR分级情况,原位杂交法检测EGFmRNA在各组的表达,HE染色观察病理组织学改变。结果:给药7d后,B,D,E组PVR分级比较,差异无统计学意义(P>0.05),给药30d后,C组与各组比较,差异有统计学意义(P<0.05);在C组,ERM中EGFmRNA阳性程度低于B组,差异有显著统计学意义(P<0.01)。D组与E组也能降低EGFmRNA表达的阳性程度,与B组相比有统计学差异(P<0.05)。C组中EGFmRNA阳性程度低于另两个治疗组,有统计学差异(P<0.05)。C组中EGFmRNA阳性程度略高于A组,有统计学差异(P<0.01)。结论:活血利水法是活血化瘀和利水明目两者作用的协同,能通过拮抗ERM中EGFmRNA表达的作用来抑制增殖细胞的过度增生,从而防治PVR形成和发展。     

散血明目片对外伤性增生性玻璃体视网膜病变兔眼血小板源性生长因子表达的影响

目的探讨散血明目片对外伤性增生性玻璃体视网膜病变（proliferative vitreoretinopathy．PVR）增殖膜上血小板源性生长因子（platelet derived growth factor,PDGF）表达的影响以及防治PVR的作用机制。方法将40只成年青紫蓝兔兔眼造成外伤性PVR模型,随机分为空白组（A组）、模型组（B组）、利水明日组（C组）、活血化瘀组（D纽）、活血利水组（E组）,每组8眼。并利用改良免疫组织化学方法对PVR实验兔眼中增殖膜上PDGF的表达进行检测。结果给药后第7天．B、C、D、E组的PVR分级比较差异无统计学意义（P〉0．05）：给药后第28天．E组与其他各组比较,差异有统计学意义（P〈O．05）。各用药组玻璃体腔增生膜PDGF阳性细胞数明显少于B组,差异有统计学意义fP〈0．05）或非常显著的统计学意义（p〈0．01）;E组与C组、D组之间比较,差异有统计学意义fP〈0．051。结论散血明目片能通过抑制玻璃体腔中PDGF对视网膜色素上皮细胞的刺激作用．从而抑制增殖细胞的过度增生．防止PVR形成和发展。     

中药散血明目片抑制兔视网膜静脉阻塞后视网膜VEGF和bFGF的表达

目的:探讨活血通脉利水明目作用的中药散血明目片防治视网膜静脉阻塞(retinal vein occlusion,RVO)并发新生血管的作用。方法:将家兔48只96眼随机分为4组,每组12只24眼。A健康空白组;B模型组;C血栓通组;D散血明目片组。采用激光光凝法建立兔RVO模型,分别于术后3,7,14,21d用空气栓塞法处死动物,即刻摘取眼球石蜡包埋切片。免疫组化法观察兔视网膜组织中血管内皮生长因子(VEGF)和碱性成纤维细胞生长因子(bFGF)的表达。结果:成功建立了RVO模型。D组与B组比较,视网膜组织中VEGF含量(21d时分别为0.442±0.034和0.583±0.056)和bFGF含量(21d时分别为0.419±0.040和0.514±0.056)明显减少,差异有统计学意义(P<0.05),与C组比较差异无统计学意义。结论:活血通脉利水明目之散血明目片能抑制视网膜面VEGF和bFGF的高表达。     

复方中药散血明目片对兔视网膜静脉阻塞ET-1表达的影响

目的:探讨活血通脉利水明目之复方中药散血明目片对兔视网膜静脉阻塞RVO模型ET-1表达的影响。方法:取健康有色家兔36只,每组9只,分为A组:健康空白组,B组:模型组,C组:血栓通组,D组:散血明目片组。采用波长532nm的Nd:YAG激光光凝法建立RVO模型,连续灌胃,实验后7,14,21d分别处死家兔取材,石蜡包埋切片,用免疫组化法检测视网膜组织中ET-1表达活性,计算机系统图像分析结果并进行统计分析。结果:D组7,14,21d与A,B,C3组比较:ET-1表达活性明显低于B组(P<0.05),与C组差异无显著性;21d时D组ET-1表达活性与A组差异无统计学意义。结论:活血通脉利水明目之复方中药散血明目片能够明显降低兔视网膜静脉阻塞RVO模型ET-1表达,抗血栓形成,改善RVO后视网膜局部微循环,减轻缺血缺氧对血管内皮细胞的损害。     

复方中药散结明目片对兔玻璃体积血所致增生性玻璃体视网膜病变的作用

目的:探讨复方中药散结明目片对玻璃体积血兔玻璃体及视网膜匀浆中IL-6含量、积血吸收及增殖膜形成的抑制作用。方法:兔自体血造成玻璃体积血模型,设空白组、模型组、和血明目片组、散结明目片高、中、低剂量组,6wk后B超观察积血及增殖膜情况,用ELISA法检测各组玻璃体及视网膜中IL-6含量。结果:B超示散结明目片高、中剂量组积血消散明显优于低剂量组及和血明目片组,未形成增殖膜。散结明目片各剂量组及和血明目片组兔玻璃体及视网膜匀浆中IL-6含量较空白组明显升高,差异有统计学意义(P<0.05),较模型组含量降低,差异有统计学意义(P<0.05)。散结明目片高、中剂量组IL-6含量明显低于低剂量组及和血明目片组,差异有统计学意义(P<0.05),与和血明目片组及模型组相比,散结明目片高、中剂量组对机化条索及增殖膜有不同程度的抑制,差异有统计学意义(P<0.05)。结论:复方中药散结明目片能促进积血吸收,抑制机化条索及增殖膜的形成。     

活血利水明目颗粒治疗超声乳化术后角膜水肿初步观察

目的:探讨活血利水明目颗粒治疗超声乳化白内障术后角膜水肿的作用。方法:将超声乳化白内障术后第1d发生2级以上角膜水肿的病例随机分为常规用药组与活血利水明目颗粒组,每组45例。常规用药组:给予妥布霉素地塞米松眼液、复方托吡卡胺眼液、重组牛碱性成纤维细胞生长因子眼液交替滴眼。活血利水明目颗粒组:在常规西药治疗的同时,加服具有活血利水、明目退翳功效的中药活血利水明目颗粒,观察两组患者的临床疗效与角膜水肿消退时间。1wk为1疗程,2个疗程结束时统计疗效。结果:活血利水明目颗粒组的临床治愈率与角膜水肿平均消退时间均明显优于常规用药组(P均<0.05),总有效率与常规用药组比较无明显差异(P>0.05)。结论:活血利水明目颗粒治疗超声乳化白内障术后角膜水肿具有较好的临床疗效,能缩短角膜水肿消退时间,提前恢复患者视力。     

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。     

活血利水法对兔视网膜静脉阻塞后视网膜抗氧化能力影响的研究

目的:探讨活血利水之散血明目片对视网膜静脉阻塞(retinal vein occlusion,RVO)后视网膜抗氧化能力的影响。方法:将40只家兔随机分为4组,每组10只兔10眼,采用光化学法建立RVO模型,4组分别给药,A组健康空白组,B,C,D组造模后分别以生理盐水、血塞通片混悬液、散血明目片混悬液灌胃。实验后7,21d处死家兔,取出视网膜检测视网膜中超氧化物歧化酶(SOD)、丙二醛(MDA)含量。结果:成功建立了RVO模型30眼,D组SOD浓度明显高于B,C两组,且差异有显著性(P<0.01);D组MDA浓度明显低于B,C两组,有显著性差异(P<0.05)。结论:活血利水之散血明目片能有效的提高视网膜抗氧化的能力,减少并发症,保护视网膜。     

增殖性糖尿病视网膜病变基质金属蛋白酶的表达

目的:研究增殖性糖尿病视网膜病变(proliferative diabetic retinopathy,PDR)视网膜增殖膜纤维化和新生血管形成与MMP-2和MMP-9异常表达的关系.方法:采用免疫组织化学方法,检测PDR患者20例视网膜前纤维血管膜MMP-2和MMP-9的表达,并与增殖性玻璃体视网膜病变(proliferative vitreoretinopathy,PVR)10例非血管性增殖膜进行对比研究.结果:PDR增殖膜MMP-2和MMP-9染色阳性率分别为70%和85%,PVR增殖膜MMP-2和MMP-9染色阳性率分别为80%和60%.在PDR增殖膜上可见血管周围基质MMP-9染色阳性,而且与PVR增殖膜相比,PDR增殖膜MMP-9表达有显著提高.结论:PDR增殖膜MMP-2和MMP-9均有表达,在PDR新生血管形成和纤维化的病理过程中起重要作用.     

益气养阴活血利水法对兔视网膜脱离复位视网膜电图的影响

目的:观察益气养阴活血利水之中药复明片对兔视网膜脱离及复位后视网膜电图的影响。方法:采用视网膜下腔注射透明质酸钠造成视网膜脱离模型(10～14d后视网膜自动复位),并于造模后1wk(视网膜脱离未复位)及造模后3wk(视网膜复位早期)对正常组、模型组、西药组、益气养阴活血利水组行视网膜电图(ERG)检查。结果:益气养阴活血利水法能在视网膜脱离时及复位后提高ERGa,b波振幅,缩短其潜时,与模型组及西药组比较有极显著性差异。结论:益气养阴活血利水法(复明片)能保护视网膜脱离视功能,并促进视网膜复位后视功能恢复。     

增生性玻璃体视网膜病变模型眼内定向 转染报告基因的研究

目的观察目的基因在玻璃体增生膜的表达情况,以探讨基因治疗增生性玻璃体视网膜病变(proliferative vitreoretinopathy,PVR) 的可行性。方法用β-半乳糖苷酶基因作为报告基因,以逆转录病毒载体携带,直接注入PVR模型眼玻璃体腔中,观察其在PVR 眼各组织表达情况。结果 基因转染后可见玻璃体增生膜组织有转染基因表达,表达主要位于增生膜的表面,而视网膜组织及其它眼组织未见表达。结论逆转录病毒载体用于PVR的基因治疗有靶向性作用,表明PVR基因治疗具有可行性。(中华眼底病杂志,2001,17:224-226)     

The effect of prinomastat (AG3340), a synthetic inhibitor of matrix metalloproteinases, on posttraumatic proliferative vitreoretinopathy

In a search for a pharmacologic adjuvant in the management of posttraumatic proliferative vitreoretinopathy (PVR), we investigated the effect of intravitreal injection of prinomastat (AG3340) on an experimental model. Posterior penetrating eye trauma was created in one eye each of 24 New Zealand white rabbits. One week after the surgery, all rabbits were randomized (1:1) to receive 0.5 mg prinomastat or the vehicle of the drug intravitreally every week for 6 weeks. The degree of PVR for each hemiretina was scored, and the two scores were summed to obtain a total eye score. The mean total eye score was 3.58 in the treatment group and 5.75 in the control group (p = 0.0307). The numbers of eyes with tractional retinal detachment in the prinomastat-treated (n = 12) and control (n = 12) groups were 3 and 9, respectively (p = 0.0391). These results suggest that intravitreally administered prinomastat has an inhibitory effect on posttraumatic PVR.     

转化生长因子-β受体Ⅱ在增生性玻璃体视网膜病变增生膜中的表达

目的:观察及评估转化生长因子-β受体Ⅱ(TGF-βRⅡ)在增生性玻璃体视网膜病变(PVR)增生膜中的表达及临床意义。 方法:采用免疫组化和原位杂交方法,对13例PVR患者行玻璃体手术增生膜获得的16例进行TGF-βRⅡ的蛋白和mRNA的检测。 结果:免疫组化结果为:9例C2～C3级膜中,染色反应为阳性的有8例,总阳性率为88.9％。7例D1～D3级膜中有6例为阳性,总阳性率为85.7％。阳性细胞多是一类胞体为长圆形,胞核呈圆形或卵圆形的上皮样细胞。统计学分析TGF-βRⅡ标记与膜分级间无相关性(P>0.05)。原位杂交结果与免疫组化基本一致。 结论:PVR发生过程中视网膜色素上皮细胞在生长因子等的刺激下,TGF-βRⅡ表达显著上调,表明了TGF-β参与PVR增生膜的形成。眼科学报 2003;19:244-247。     

p21WAF1/CIP1在兔外伤性增生性玻璃体视网膜病变发生和发展中的抑制作用

背景 p21是一种细胞周期蛋白依赖性激酶抑制剂,能阻止细胞从G1期进入S期,抑制细胞增生,研究认为内源性p21表达的动态变化可能与细胞增生性病变有关.外伤性增生性玻璃体视网膜病变(PVR)是眼部增生性反应相关性疾病,了解PVR过程中p21表达的动态变化可能为PVR的靶向治疗提供依据.目的 检测p21WAFl/CIP1在兔外伤性PVR中的动态变化,探讨其在外伤性PVR发病机制中的作用.方法 选取青紫蓝兔54只,采用随机数字表法将实验兔随机分为正常对照组(6只)和造模后7、14、21和28 d组(每组12只),每只兔任意选取一眼作为实验眼.各模型组兔眼玻璃体腔注射人富含血小板血浆(PRP)0.4 ml,同时于鼻上方角巩膜缘后5 mm处行巩膜外冷冻约5 s,以建立外伤性PVR模型.各组兔眼行眼部B型超声检查以评估建模情况.分别于造模后7、14、21和28 d以过量麻醉法处死实验兔并制备实验眼视网膜组织切片,采用苏木精-伊红染色法检测兔眼视网膜的形态表现,分别采用免疫组织化学染色、Western blot及逆转录PCR(RT-PCR)法检测兔视网膜中p21WAFl/CIP1蛋白及其mRNA的相对表达. 结果 正常对照组兔眼眼前后节均正常,模型组兔造模后1～7 d兔眼玻璃体中增生条索逐渐变粗,可见视网膜皱褶,造模后14d兔眼出现牵引性视网膜脱离,造模后28 d兔眼漏斗状视网膜脱离.视网膜病理组织学检查显示,造模后7d兔眼视网膜表面有增生膜和炎性细胞聚集,造模后28 d可见视网膜呈花瓣形固定皱褶,视网膜结构紊乱.免疫组织化学染色显示,p21WAF1/CIP1蛋白在正常对照组兔眼视网膜神经节细胞层及内核层的细胞核内呈强阳性表达,造模后7、14、21和28 d表达强度减弱,以造模后14d表达量最低.Western blot结果显示,正常对照组和造模后7、14、21和28 d组兔眼视网膜中p21WAF1/CIP1蛋白相对表达量分别为0.74±0.08、0.60±0.05、0.56±0.03、0.74±0.02和0.65±0.04,组间总体比较差异有统计学意义(F=20.55,P=0.00),造模后7d和14d的表达量均明显低于正常对照组和造模后21 d和28 d组,差异均有统计学意义(均P＜0.05).RT-PCR结果显示,正常对照组和造模后7、14、21和28 d组兔眼视网膜中p21WAF/CIP1 mRNA的相对表达量分别为0.65±0.09、0.57±0.05、0.45±0.04、0.46±0.02和0.47±0.04,总体比较差异有统计学意义(F=18.06,P=0.00),造模后14、21和28 d P21WAF1/C1P1 mRNA的相对表达量均明显低于正常对照组和造模后7d组,差异均有统计学意义(均P＜0.05).结论 p21WAF1/CIP1在外伤性PVR兔眼视网膜中的动态表达变化与PVR的病程发展过程相吻合,p21可能参与外伤性PVR发生和发展的病理过程,p21WAF1/C1P1表达量的下降与细胞增生的动态变化过程一致,可能促进了PVR的进展.     

The effect of prinomastat (AG3340), a potent inhibitor of matrix metalloproteinases, on a subacute model of proliferative vitreoretinopathy

PURPOSE. To determine the efficacy of prinomastat (AG3340), a synthetic inhibitor of matrix metalloproteinase, in the treatment of experimental proliferative vitreoretinopathy (PVR) induced by intravitreal dispase injection. METHODS. One eye each of 53 New Zealand white rabbits was injected in the vitreous cavity with 0.07 unit of dispase to induce PVR. One week after PVR induction, 53 rabbits were randomized (27:26) to receive 0.5 mg prinomastat or the vehicle of the drug (acidified water) intravitreally every two weeks. The scores of PVR severity (scale of 1-5) were graded to compare the prinomastat-treated animals with the control group. RESULTS. The average PVR scores in the treatment and control groups were 2.62 and 3.57 respectively (p = 0.038; Wilcoxon rank sum). Clinically significant PVR with retinal detachment (PVR > or = grade 3) developed in 76% of rabbits in the control group versus 51% of rabbits treated with prinomastat. CONCLUSIONS. Intravitreally administered prinomastat decreased development of PVR in an experimental model which made use of dispase to induce PVR.     

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）     

The effect of tranilast on experimental proliferative vitreoretinopathy

· Background: Tranilast has been clinically used for various allergic diseases. Recently, it has also been found to inhibit excessive scarring in wound healing processes. In this study, we examined the effects of tranilast on the treatment for experimental proliferative vitreoretinopathy (PVR). · Methods: Cultured rabbit conjunctival fibroblasts were injected intravitreously (50 000 cells/eye) into the rabbit vitreous to induce experimental PVR. Immediately after that, tranilast (0.5–5 mg/ml, 0.1 ml/eye) was injected into the vitreous. Injection of vehicle solution was used as a negative control. PVR was clinically evaluated by masked observers using ophthalmoscopy and graded into six stages: 0 (no PVR) to 5 (severe PVR). The amount of transforming growth factor β1 (TGF-β1) in the vitreous was measured by ELISA method. Functional and morphological changes induced by 5 mg/ml tranilast were sought by electroretinography, light microscopy, and electron microscopy on day 28. · Results: The average stage of PVR in the eyes treated with tranilast (1 or 5 mg/ml) was significantly lower than that in the control group on days 14 and 28. There was no difference between the eyes treated with low-dose tranilast (0.5 mg/ml) and the control group. The amount of TGF-β1 in the vitreous of tranilast-treated eyes was significantly lower than in the control group. The morphological and functional studies did not show any deleterious effect of tranilast on the retinal function and morphology. · Conclusion: Tranilast effectively inhibits the progression of PVR without showing apparent toxicity of the eye. This agent has therapeutic value for PVR. Received: 4 November 1998 Revised version received: 18 January 1999 Accepted: 1 February 1999     

Soluble TNF receptors in vitreoretinal proliferative disease

PURPOSE: To measure vitreous levels of soluble TNF-receptors (sTNF-Rs) types I and II in eyes with rhegmatogenous retinal detachment (RRD), uncomplicated or complicated with proliferative vitreoretinopathy (PVR), and in eyes with proliferative diabetic retinopathy (PDR). To examine whether there is any relationship between vitreous levels of sTNF-Rs and clinical features of these conditions and between vitreous sTNF-Rs and TNFalpha levels and serum levels of sTNF-RS: METHODS: Vitreous levels of sTNF-Rs and TNFalpha were measured by enzyme-linked immunosorbent assay in 30 eyes with PVR, 30 eyes with uncomplicated RRD, and 29 eyes with PDR. Vitreous from eyes of 10 deceased donors and 9 eyes with macular holes served as control specimens. Serum levels of sTNF-Rs were measured in 17 patients with PDR and 21 patients with PVR. RESULTS: Vitreous levels of sTNF-Rs I and II were increased in eyes with PVR, RRD, and PDR when compared with control eyes (P < 0.002). However, vitreous levels of sTNF-Rs I and II were higher in eyes with PVR than in eyes with RRD (P < 0.01) or PDR (P < 0.03). This contrasted with the findings that serum sTNF-Rs were higher in PDR than in PVR (P < 0.016) and that vitreous levels of TNFalpha were higher in eyes with PDR than in eyes with PVR (P < 0.0005). In PVR, vitreous sTNF-Rs levels were associated with the duration of retinal detachment, number of previous external operations, and grade of severity, whereas in PDR these levels were not related to the type or duration of diabetes or its complication with traction retinal detachment. CONCLUSIONS: These observations suggest the existence of TNF inhibitory mechanisms within the eye during retinal processes of inflammation and angiogenesis. That high vitreous levels of sTNF-Rs relate to severity of retinopathy suggests that these molecules may constitute reactive products of inflammation. Effective control of TNFalpha activity by sTNF-Rs within the retinal microenvironment may determine the outcome and severity of retinal proliferative conditions.