柔毛霉素对兔晶体上皮细胞体外生长抑制的实验研究

目的：研究柔毛霉素（ｄａｕｎｏｍｙｃｉｎ）对兔晶体上皮细胞体外生长抑制，探索使用柔毛霉素以预防后发障的发生。方法：分离培养家兔晶体上皮细胞，传代后在２４孔培养板培养７２小时，计数后加入不同浓度的柔毛霉素作用１０分钟后，吸出药物并冲洗后培养５天，结果：柔毛霉素的ＬＤ５０为０．５μｇ／ｍｌ～０．７７μｇ／ｍｌ，经高浓度药物（１０μｇ／ｍｌ）作用后的标本几乎无细胞存活；经０．５μｇ／ｍｌ药物浓度作用后的细胞形态发生变化。结论：显示出低浓度柔毛霉素短时间作用后即能有效抑制晶体上皮细胞的增生。     

MTT比色法测定药物对晶体上皮细胞增殖的影响

目的筛选防治后囊混浊的药物。方法进行人胚胎晶体上皮细胞（ｌｅｎｓｅｐｉｔｈｅｌｉａｌｃｅｌｓ，ＬＥＣ）的培养并用ＭＴＴ比色法检测了肝素、氟脲嘧啶、双氯灭痛、顺铂对人胚胎晶体上皮细胞增殖的影响。结果肝素１００～２５００Ｕ／ｍｌ，氟脲嘧啶０．１～１００μｇ／ｍｌ，顺铂１０～５００μｇ／ｍｌ作用２４ｈ和７２ｈ对胚胎晶体上皮细胞增殖有明显抑制作用（Ｐ＜０．０５）。作用７２ｈ抑制率高于２４ｈ（Ｐ＜０．０５）。双氯灭痛和对照组相比无明显差异（Ｐ＞０．０５）。结论肝素、氟脲嘧啶、顺铂均为剂量依赖型和时间依赖型药物，均能有效抑制晶体上皮细胞的生长；双氯灭痛对晶体上皮细胞增殖无明显抑制作用。     

阿霉素,5—Fu,三尖杉醌碱及去炎松对体外培养的人视网膜色素上皮细胞…

应用＾３Ｈ－胸腺嘧啶核苷掺入及液体闪烁测量技术，观察不同浓度的阿霉素，５－氟尿嘧啶，三尖杉酯碱及去炎松对体外培养的人视网膜色素上皮细胞增殖的抑制作用。结果；阿霉素在２－３２ｎｍ／ｍｌ时，对ＲＰＥ细胞抑制率为８．５－７７．２＾，ＩＤ５０为１２ｎｇ／ｍｌ；５－Ｆｕ在０．２－３．２μｇ／ｍｌ时，对ＲＰＥ细胞抑制率为１４．５－８１．７％，ＩＤ５０为０．６８μｇ／ｍｌ；三尖杉酯碱在２．５－４０ｎｇ／ｍｌ，抑     

表皮生长因子对晶体上皮细胞增殖影响的研究

目的 评价表皮生长因子对晶体上皮细胞增殖的影响。方法 在培养的牛和人晶体上皮细胞中添加ＥＧＦ,甲基噻唑基四唑法测定细胞增殖情况以及一抗体对ＥＧＦ作用的影响。结果ＥＧＦ浓度为１０＾－１－１０＾２μｇ／Ｌ对牛晶体上皮细胞增殖有促进作用,其中浓度为１０μｇ／Ｌ作用３天达到最大促增殖效果;ＥＧＦ浓度为１－１０＾２μｇ／Ｌ对人晶体上皮细胞增殖有促进作用,浓度为１０＾２μｇ／Ｌ具有最大促增殖效果。抗ＥＧＦＲ抗     

地塞米松和柔红霉素抑制后囊混浊的体外模拟研究

为评价地塞米松和柔红霉素防治后囊混浊的价值，在体外模拟其在体内的可行性用药方式观察其对牛晶体上皮细胞增殖的抑制作用，结果地塞米松模拟在术后用药７２小时，在房水有效浓度内对晶体上皮细胞无明显抑制作用；提示其防治后囊混浊的作用主要是通过加速血－房水屏障的恢复，减少和控制术后炎症反应而间接实现。柔红霉素模拟加入皮质冲洗液中作用１０分钟，在眼部允许剂量内可呈浓度依赖性抑制晶体上皮细胞增殖，认为按这种方式用药可达到抑制后囊混浊作用。     

维拉帕米,肝素,地塞米松抑制后囊混浊的细胞学基础：对晶体上皮细胞…

通过观察维拉帕米，肝素，地塞米松在体外对牛晶体上皮细胞（ＢＬＥＣ）增殖和细胞内Ｃａ＾＋的影响评价一它们抑制后囊混浊的价值。结果维拉帕米呈浓度依赖性抑制ＢＬＥＣ增殖（Ｐ〈０．０５（１０ｍｇ／Ｌ）或Ｐ〈０．０１（≥２０ｍｇ／Ｌ）和降低细胞内Ｃａ＾＋并能增强肝素的增殖抑制作用肝素５００ｍｇ／Ｌ时可明显抑制增殖（Ｐ〈０．０１）和降低细胞内Ｃａ＾＋＋５００ｍｇ／Ｌ以上抑制作用不随浓度增加而增强，地塞米松对Ｂ     

抗增生药物对人视网膜神经胶质细胞的抑制作用

目的寻找抑制视网膜、玻璃体内细胞增生的有效药物;明确抗增生药物秋水仙碱、道诺霉素和５－氟尿嘧啶（５－Ｆｕ）对体外培养的人视网膜胶质（ｒｅｔｉｎａｌ ｇｌｉａ,ＲＧ）细胞的作用。方法用 ＭＴＴ法测定秋水仙碱（０．５～１６．０μｇ／ｍｌ）、道诺霉素（０．１～３．２μｇ／ｍｌ）和５－Ｆｕ（０．５～１６．０μｇ／ｍｌ）对体外培养人ＲＧ细胞的作用。结果秋水仙碱（１．０～１６．０μｇ／ｍｌ）、道诺霉素（０．２～３．２０μｇ／ｍｌ）和 ５－Ｆｕ（１．０～１６．０μｇ／ｍｌ）３组药物对培养细胞均有抑制作用,与对照组均差别显著（Ｐ＜０．０１）,ＩＤ_（５０）分别为３．１１μｇ／ｍｌ,０．７９μｇ／ｍｌ和５．２３μｇ／ｍｌ。结论秋水仙碱、道诺霉素和５－Ｆｕ对ＲＧ细胞有明显抑制作用。     

晶体上皮细胞在体外的分化规律及血清和眼组织对其增殖的影响

目的从细胞培养水平探讨后囊混浊形成机理和术后血－房水屏障破坏、晶体皮质残留等对其的影响。方法用考马斯亮蓝（ＣｏｏｍａｓｓｉｅＢＢ）染色、倒置显微镜和电镜观察培养的牛晶体上皮细胞（ｂｏｖｉｎｅｌｅｎｓｅｐｉｔｈｅｌｉａｌｃｅｌｓ，ＢＬＥＣ）的增殖和分化规律，用Ｇｉｅｍｓａ染色比色法观察胎牛血清、房水、晶体皮质等对ＢＬＥＣ增殖的影响。结果在体外培养条件下ＢＬＥＣ可在第１～７代内分化为晶体纤维细胞，表现为细胞体积增大，形状渐趋长条形和梭形，细胞骨架逐渐增多；胎牛血清呈浓度依赖性促进ＢＬＥＣ增殖（Ｐ＜０．０５）；高浓度房水（占培养液１／３）抑制ＢＬＥＣ增殖（Ｐ＜０．０１），晶体皮质、晶体核、玻璃体均促进ＢＬＥＣ增殖（Ｐ＜０．０１）。结论晶体上皮细胞的分化在后囊混浊形成中起重要作用；术后血－房水屏障破坏、晶体皮质残留和玻璃体脱出可通过刺激晶体上皮细胞增殖促进后囊混浊形成。     

生长抑素对体外培养的人晶体上皮细胞的增殖和胶原合成的抑制作用

目的研究生长抑素ｓｔｉｌａｍｉｎ（ＳＳ－１４肽）和ｓａｎｄｏｓｔａｔｉｎ（ＳＳ－８肽）对人晶上皮细胞（ＨＬＥＣ）增殖及胶原合成的影响。方法采用ＭＴＴ比色法和３Ｈ－脯氨酸（３Ｈ－Ｐｒｏｌｉｎｅ）掺入法检测了ＳＳ－１４肽和ＳＳ－８肽对人晶体上皮细胞（ＨＬＥＣ）增殖及胶原合成的抑制作用。结果ＳＳ－８肽浓度为１０－１０ｍｏｌ／Ｌ、ＳＳ－１４肽浓度为１０－９ｍｏｌ／Ｌ时能明显抑制ＨＬＥＣ增殖和胶原合成，且呈剂量依赖关系，５０％抑制浓度（ＩＤ５０）ＳＳ－８肽为１０－８ｍｏｌ／Ｌ，ＳＳ－１４肽为１０－７ｍｏｌ／Ｌ。结论ＳＳ－１４肽和ＳＳ－８肽可以通过抑制ＨＬＥＣ的增殖及胶原的合成而抑制后囊混浊。     

肿瘤坏死因子α和表皮生长因子对晶状体上皮细胞增殖的作用

目的研究肿瘤坏死因子α（ＴＮＦα）和表皮生长因子（ＥＧＦ）对人、牛晶状体上皮细胞增殖的影响。方法采用ＭＴＴ比色法检测ＴＮＦα和ＥＧＦ对晶状体上皮细胞（ｌｅｎｓｅｐｉｔｈｅｌｉａｌｃｅｌ，ＬＥＣ）增殖的作用。结果０．１Ｕ／ｍｌ的ＴＮＦα即可明显促进人、牛晶状体上皮细胞的增殖；ＥＧＦ为１、１０ｎｇ／ｍｌ时可明显促进牛晶状体上皮细胞增殖。结论细胞因子ＴＮＦα、ＥＧＦ通过促进ＬＥＣ的增殖参与后囊混浊的形成     

柔红霉素对人晶体上皮细胞生长抑制和角膜内皮细胞毒性试验研究

目的：研究柔红霉素对体外培养人晶体上皮细胞增殖的抑制作用和对角膜内皮细胞的毒性，探索使用柔红霉素预防后发障的适宜浓度，方法：第三代人晶 皮细胞接种于24孔培养板内，24h后加入不同浓度柔红霉素作用10min后，吸出药物并冲洗后再培养48h，细胞计数，原代培养角加入膜内皮细胞，加入不同浓度柔红霉素作用10min，观察细胞生长形态学变化，结果：柔红霉素依浓度梯度抑制晶体上皮细胞增殖，其LD50为4．9ug/ml，10ug/ml以下浓度对角膜内皮细胞无损害，结论：柔红霉素有可能作为预防后发障的辅助药物之一，选择适宜浓度是防止其对其他组织产生毒性的关键。     

晶体囊袋内应用丝裂霉素防治兔后发性白内障的实验研究

目的：探索晶体囊代内灌注丝裂霉素对兔晶体上皮细胞的抑制作用及对兔眼的毒性作用。方法：在兔晶状体超声乳化吸出术中,用0．2ml不同浓度的丝裂霉素（Mitomycin C,MMC)(0.1mg/ml、0.2mg/ml、0.4mg/ml）在晶体囊代内进行水分离,使其直接短暂作用于晶体上皮细胞。术后随访2个月,观察比较用药组和对照组兔晶体后囊混浊、眼内压的变化等;并观察术后组织病理学和超微结构的变化。结果：临床观察显示,术后2周只有对照眼出现后囊膜混浊（posterior capsule opacification,PCO)。术后4周用药组眼的PCO明显比对照组眼轻,且随药物浓度增高而减轻。中、低浓度组兔眼的术后炎症反应与对照组眼比较无明显差异。高浓度组兔眼术后早期出现轻微的毒性反应。组织,病理学检查表明,对照组的晶体上皮增生较用药组明显。MMC可引起晶体上皮细胞变性,其变化程度与药物浓度有关。高浓度眼睫状体有炎症细胞浸润及少量出血。结论：MMC有效地抑制晶体上皮细胞的增殖,其作用强度与药物的浓度相关。兔晶体囊贷内应用0．1－0．2mg/ml的药物浓度是防治后发性白内障安全有效的药物。     

体外模拟牛眼晶状体后囊膜混浊及普拉洛芬眼液对其细胞融合影响的实验研究

目的探讨晶状体后囊膜上晶状体上皮细胞的增生分化规律和普拉洛芬滴眼液对晶状体后囊膜混浊（PCO）的抑制作用.方法将牛后囊膜晶状体上皮细胞面朝上平铺于25 ml培养瓶底部,分别加入含0%、10%、20%胎牛血清的DMEM培养液培养1～5周,计数后囊膜晶状体上皮细胞覆盖率,并用Giemsa染色和扫描电镜观察其形态.同时用相当于普拉洛芬滴眼液滴眼后4 h在房水中的浓度（0.23 mg/L）作用于组织培养模型,与对照组比较,观察其对后囊膜晶状体上皮细胞融合时间的影响.结果赤道部的晶状体上皮细胞向晶状体后囊膜增生、迁徙,晶状体后囊膜形成明显的混浊和皱缩;后囊膜晶状体上皮细胞融合天数随血清浓度的升高而缩短;浓度为0.23 mg/L的普拉洛芬对后囊膜晶状体上皮细胞的融合有明显抑制作用.讨论体外模拟PCO的组织培养是研究后发性白内障的有效方法.普拉洛芬是一种安全、有效降低PCO发生率的药物,可作为白内障术后的常规用药.     

Retrovirus-mediated transfer of a suicide gene into lens epithelial cells in vitro and in an experimental model of posterior capsule opacification.

PURPOSE. The most common complication of cataract surgery is the development of posterior capsule opacification (PCO). Hyperplasia of the lens epithelium is one of the main cellular events following phacoemulsification and was found to be an important feature contributing to opacification of the posterior capsule. We investigated the feasibility of killing the residual lens epithelial cells by retroviral-mediated transfer of the herpes simplex virus-thymidine kinase (HSV-tk) gene, a well-studied suicide gene, into rabbit lens epithelial cells followed by ganciclovir (GCV) treatment. METHODS. The capacity of retroviral vectors to transfer genes into rabbit lens epithelial cells was determined either in vitro (culture of rabbit lens epithelial cells) or in vivo (experimental model of PCO in rabbits) using cDNA encoding the beta-galactosidase (LacZ) reporter gene. To evaluate the efficiency of suicide gene therapy (infection with retroviral vectors encoding the HSV-tk gene followed by GCV treatment) we determined the sensitivity of HSV-tk infected lens epithelial cells to different concentrations of GCV in vitro. Then, in an experimental model of PCO, rabbits were treated with HSV-tk retroviral vectors at the end of the surgery and they received repeated intracameral and intravitreal injections of GCV at the concentration determined by the in vitro experiments. RESULTS. Infection efficiency using LacZ retroviral vectors was about 29% in vitro and 10% in vivo. After infection of the HSV-tk cDNA in vitro, the cell killing effect of GCV was evaluated. A significant enhancement (four- to five-fold) of the cell sensitivity to GCV was shown in FLY-DFGtk as compared with mock infected (P < 0.01) cells even without selection of the HSV-tk positive cells. The GCV concentration leading to 50% reduction in cell number (IC50) was 50 microg/ml. In vivo infection with a HSV-tk vector led to the tk gene transfer into lens epithelial cells. Despite this local HSV-tk gene expression, we could not prevent capsule opacification. CONCLUSIONS. Lens epithelial cells were successfully infected both in vitro and in vivo by beta-galactosidase and HSV-tk genes via retroviral vectors. In vitro infected lens epithelial cells displayed a strong sensitivity to GCV treatment. In vivo, we could not prevent capsule opacification in the rabbit model, very likely due to the limited level of the HSV-tk gene expression. However, our results suggest that virus-mediated suicide gene therapy might be a feasible treatment strategy to prevent capsule opacification with a more powerful vector.     

Evaluation of daunomycin toxicity on lens epithelium in vitro

Posterior capsule opacification is the major complication of extracapsular cataract extraction (ECCE). Lens epithelial cells derived from the periphery of the lens are thought to migrate posteriorly and contribute significantly to the postoperative proliferations at the posterior pole. We have evaluated the effects of the antiproliferative drug daunomycin on cultured porcine lens epithelial cell viability and proliferation. We observed that the mitotic activity of the cells is suppressed by a single short time treatment with daunomycin at a concentration as low as 2.5 mg/l. Long term effects on the reproductive capacity of the lens epithelial cells may not be as pronounced as the inhibition of other cells examined before e.g. retinal pigment epithelium and fibroblasts. Our results indicate that daunomycin may be useful for the pharmacologic prevention of postoperative proliferations in patients treated by ECCE.This study was supported by the Retinovit-Foundation.     

The Effect of Liposome Encapsulated Daunorubicin on Rabbit Eyes after Extracapsular Lens Extraction

Purpose: To investigate the effect of liposome encapsulated daunorubicin (DNR) on rabbit eyes when it was used in prevention of posterior capsule opacification (PCO). Methods: The liposome encapsulated DNR was prepared by modified freeze-thawing method. Each eye was injected with 0. 1 ml liposomes (0. 2 mg/ml and 20 μg/ml DNR) into the capsular bag during the extracapsular lens extraction (ECLE) in 10 rabbit eyes respectively. The phosphate buffer solution (PBS) was injected as control. Besides biomicroscope observation and histology examination of all eyes, the concentration of DNR in aqueous humor was also determined by high performance liquid chromatography (HPLC).Results: The morphology of liposome encapsulated DNR were similar to the blank liposome with round or ellipse shape. The encapsulated effeciency of liposome encapsulated DNR was 45. 1%. The inflammatory response was much more severe both in 0. 2 mg/ml and 20μg/ml DNR group than the control after liposome injection. All eyes in DNR group w     

靶向治疗后囊混浊的研究进展

后囊混浊是白内障手术的常见并发症。残留晶状体上皮细胞在后囊的增殖、移行、纤维化生是后囊混浊形成的重要因素。用于防治后囊混浊的许多约物如道诺霉素、丝裂霉素等，因眼内的毒副作用而受到限制。靶向治疗的特异性、针对性为解决这一问题带来了希塑。综述了靶向治疗后囊混浊的研究现状及对未来的研究展望。     

Cyclosporin effectively inhibits posterior capsule opacification after phacoemulsification in rabbits: a preliminary study

Purpose: To evaluate whether cyclosporin A prevents or reduce posterior capsule opacification after phacoemulsification surgery in rabbit eyes. Methods: Twenty rabbits underwent cataract surgery in their right eyes were randomized into two groups. In group 1, 0.1 mL cyclosporin A (0.5 mg/mL) was given into the capsular bag after phacoemulsification and 0.1 mL cyclosporin A (0.5 mg/mL) was injected subconjunctivally once every 3 days for 1 week. Group 2 served as a control group. The development of posterior capsule opacification was assessed weekly, its density was graded clinically, and the proliferation of lens epithelial cells was evaluated histologically. Results: On clinical assessment, cyclosporin A was significantly effective in preventing posterior capsule opacification compared with controls (P = 0.0045). On histological analysis, there was significantly reduced proliferative activity on posterior capsules in the treatment group, in contrast to multilayer lens epithelial cells proliferation in the control group. Conclusion: The preliminary results indicate that cyclosporin A is effective in suppressing posterior capsule opacification in rabbits, yet requires further investigation.     

Lens epithelial inhibition by PMMA optic: implications for lens design

It has been a clinical impression that posterior chamber lens implants in some way inhibit opacification of the posterior lens capsule after extracapsular cataract extraction. The mechanism of this inhibition is unclear; it may be related to mechanical contact or blockage of migration of lens epithelial cells, or possibly to the leeching of toxic factors from the lens itself. A better understanding of the exact mechanism of opacification inhibition may have important clinical implications for intraocular lens design. For example, some lens designs that facilitate Nd:YAG capsulotomy by physically separating the posterior chamber lens and the posterior capsule may result in less inhibition and in fact more opacification of posterior capsules. We performed in vitro tissue culture studies of the effect of the polymethylmethacrylate (PMMA) optic of a planoconvex intraocular lens on cultured rabbit lens epithelium. These studies demonstrated both inhibition of lens epithelial migration beneath the PMMA optic (plano side down) as well as metaplasia and necrosis of lens cells growing directly beneath the optic. The clinical implications of these studies for intraocular lens design are discussed.