大鼠晶状体摘出术后LECs转分化观察

目的 观察大鼠晶状体摘出术后晶状体上皮细胞(lens epithelial cells,LECs)的形态、分布改变及其转分化的动态过程.方法 对50只SD大鼠左眼行晶状体囊外摘出术(extracapsular lens extraction,ECLE).分别于术后0 h、3 d、7 d、14 d及28 d对术眼进行裂隙灯检查并处死动物各10只,摘除眼球行光镜及免疫组化检测,Western Blot法检测各时间点后囊膜α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)表达.结果 术后0 h于前囊下和赤道部的内表面观察到LECs,后囊膜无细胞分布.晶状体后囊膜混浊在术后3 d出现,囊膜皱缩,整个晶状体囊袋内出现纺锤形细胞.7 d时后囊膜明显混浊,囊袋内见较多纺锤形细胞分布.术后14 d时出现明显后囊膜皱缩,见新生晶状体纤维,纺锤形细胞明显减少,后囊膜仍可见细胞.28 d可见明显后囊膜增厚,新生晶状体纤维填充囊袋,后囊膜未见细胞.免疫组化检测见术后3 d α-SMA阳性表达.Western Blot检测α-SMA于术后0 h几乎检测不到,3 d表达明显增强,7 d达高峰,14 d下降,28 d明显下降.结论 大鼠ECLE术后囊袋内LECs形态及分布呈动态变化,后囊膜α-SMA表达提示ECLE术后3～7 d是LECs转分化的关键时期.     

大鼠晶状体囊外摘出术后LECs基因差异表达的研究

目的 探讨采用基因芯片技术检测大鼠晶状体囊外摘出术(ECLE)后晶状体上皮细胞(LECs)基因表达的差异.方法 20只SD大鼠随机分成4组,双眼行ECLE;分别于手术后即刻及1、7、14 d处死大鼠,完整取出囊膜;用含有5 705个大鼠基因探针的芯片分别检测术后1、7、14 d与术后即刻组相比较的基因表达差异;实时荧光定量聚合酶链式反应(qRT-PCR)验证芯片检测结果 .结果 所有大鼠均成功施行双眼ECLE手术,术后出现PCO及晶状体物质再生.芯片实验质量控制可靠,结果 可信.术后3个时间点差异表达基因共694条.发生差异表达的基因范围非常广泛,涉及所有细胞功能类别.结论 成功地建立了大鼠ECLE术后PCO及晶状体物质再生模型.大鼠ECLE术后LECs基因差异表达极其复杂,提示PCO及晶状体再生的分子机制复杂,调控因素多.     

大鼠后发性白内障发生过程中晶状体纤维分化的初步研究

目的在组织学和mRNA水平上初步了解大鼠后发性白内障模型形成过程中晶状体纤维的分化。方法对48只SD大鼠行晶状体囊外摘除术,在术后即刻、1d、3d、1周、2周、1个月、2个月、3个月行裂隙灯显微镜观察和HE染色;取不同时间点后发性白内障组织,采用半定量逆转录聚合酶链反应法（RT-PCR）检测晶状体蛋白基因-αA、αB、βB1、βB2、βA2、γD的表达水平。结果所有大鼠术后均发生晶状体后囊膜混浊（PCO）。术后即刻,晶状体前囊膜下残留晶状体上皮细胞（LEC）;术后1d,LEC已增殖迁移至晶状体后囊膜中央区;术后3d,晶状体后囊膜中央轻度混浊、皱缩,晶状体囊袋内布满增生的LEC,周边部发生晶状体纤维分化。术后7d,后囊膜中央混浊继续加重,呈放射状皱褶,周边形成Seommering环。术后14d,瞳孔区囊膜组织纤维机化,Seommering环更为明显;术后1个月。新生晶状体纤维填满晶状体囊袋,形成类似正常晶状体的赤道部。术后2、3个月,晶状体纤维继续增生,体积接近正常晶状体。半定量RT-PCR显示术后αA、αB、βB1、βB2、βA2、γD mRNA的表达逐渐增加。结论SD大鼠行晶状体囊外摘除术后短期内即会发生明显的后发性白内障,并表达α、β和γ晶状体蛋白。该动物模型可用于后发性白内障的发病机制和晶状体再生的应用基础研究。     

兔眼晶状体再生模型的建立及观察

目的:建立兔眼晶状体再生模型,探讨晶状体上皮细胞生长及晶状体再生的机制。方法:在8只新西兰白兔兔眼上行超声乳化晶状体吸除术,建立晶状体囊袋模型,术后裂隙灯观察晶状体再生并行再生晶状体的组织学检查。结果:术中撕囊口较小并有相对完整囊袋结构的实验兔眼可观察到晶状体再生;再生晶状体周边及前囊下有较好的透明性,越接近中央及后囊越混浊;组织学研究表明再生晶状体的赤道部晶状体上皮细胞增殖分化形成晶状体纤维,中央区LEC未参与增殖分化;再生晶状体的混浊部位其晶状体纤维排列混乱;囊膜皱折区有大量细胞增殖。结论:保留晶状体囊袋空间结构的相对完整性,可使兔眼晶状体再生;晶状体再生是赤道部晶状体上皮细胞增殖分化形成,囊膜形态结构影响细胞的增殖和分化。眼科学报 2002;18:230-234,248.     

BALB/c小鼠后发性白内障动物模型的建立和观察

目的建立BALB／c小鼠后发性白内障（posterior capsule opacification，PCO）动物模型并检测Sox1／2胚胎晶状体发育调控基因在PCO中的表达。方法腹腔麻醉联合表面麻醉下对30只BALB／c小鼠行右眼晶状体囊外摘出术，分别于术后即刻、3d、1周、2周和1个月对术眼进行裂隙灯显微镜及组织病理学检查，观察PCO形成的时间、部位、发展过程及组织形态学改变；采用逆转录聚合酶链反应（RT-PCR）方法检测Sox1／2胚胎晶状体发育调控基因在术后不同时间点PCO中的表达。结果裂隙灯显微镜观察：后囊膜皱褶、混浊由周边部向中央区发展伴Elschnig小体和晶状体纤维生成，其程度随时间推移日渐加重；再生晶状体形态和大小与正常晶状体相似但透明度明显下降。组织病理学检查：手术后即刻，赤道部和前囊膜下可见单层晶状体上皮细胞（lens elial cell，LEC），后囊膜表面无LEc及晶状体皮质残留；术后3d，赤道部LEC增生并迁移至后囊膜。囊袋周边部LEC开始早期纤维分化，但核仍靠近后囊膜表面；术后1周，赤道部LEC继续分化，细胞伸长呈带状伴核远离后囊膜表面；术后2周，周边部晶状体纤维细胞持续增多，形成与正常晶状体赤道部形态类似的弓形带；术后1个月。新生晶状体纤维几乎填充整个残余囊袋，排列欠规则，细胞核罕见。RT-PcR检测：术后3d、1周、2周及1个月的PC0组织中可检测到Sox1／2条带；术后即刻囊袋组织中无Sox1／2表达。结论BALB／c小鼠可成功建立PCO动物模型并检测到Sox1／2胚胎晶状体发育调控基因的表达，为在分子生物学水平上进一步探索PCO的发病机制提供了有利条件，具有重要的应用价值。【眼科新进展2007；27（2）：91-95]     

改良大鼠后囊膜混浊模型的建立及晶状体上皮细胞的变化

目的:改良法建立大鼠后发性白内障动物模型并观察LEC在PCO过程中的动态变化。方法:SD大鼠50只在连续环形撕囊、水分离后行晶状体囊外摘除术(ECLE),娩核后使用无菌空气恢复前房。分别于术后0,3,7,14及28d对术眼进行裂隙灯检查并处死动物,摘除眼球行光镜观察,免疫组化检测LEC的α-SMA表达。结果:100%术眼后囊膜存在,84%的鼠眼可用于检测。术后0h于前囊下和赤道部的内表面观察到LEC。PCO在术后3d出现,出现囊膜皱缩,整个晶体囊膜出现纺锤形细胞。术后7d时后囊膜明显混浊,见较多纺锤形细胞分布。所有动物于术后14d出现明显后囊膜皱缩,可见新生晶体纤维。术后28d见明显后囊膜增厚,新生晶体纤维填充囊袋,后囊膜未见细胞。LEC形态及分布恢复到术后0h状态。免疫组化检测见术后3d时α-SMA阳性表达。结论:改良法成功建立大鼠PCO模型,LEC在PCO形成中出现形态和分布上的动态变化。其将为在分子水平上探索PCO的发病机制及防治方法提供合适研究载体。     

晶状体囊膜抛光联合囊袋内注射透明质酸酶预防兔晶状体后囊膜混浊的实验研究

晶状体后囊膜混浊(posterior capsule opacification，PCO)是白内障摘除术后最常见也是严重影响视力的并发症之一，主要是由术后残留的晶状体上皮细胞(lens epithelial cells，LECs)增殖、移行、化生以及术后炎性反应引起。我们采用晶状体囊袋内注射透明质酸酶(hyaluronidme，HS)联合囊膜抛光的方法，在44只兔眼的研究中证实它能有效降低PCO的发生，而且眼内应用安全，现报告如下。     

去整合素kistrin对兔眼晶状体后囊Ⅳ型胶原表达的影响

背景 白内障囊外摘出术后残留的晶状体上皮细胞(LECs)增生是后囊膜胶原产生进而形成后发性白内障的生物学基础,去整合素可与细胞外基质(ECM)竞争结合整合素分子,理论上可以防治后发性白内障的形成,但其具体的作用机制有待进一步研究. 目的 研究去整合素kistrin对兔眼晶状体后囊Ⅳ型胶原表达的影响.方法24只新西兰大白兔按照随机数字表法分为kistrin注射组及生理盐水对照组,每组12只.两组兔均行右眼透明晶状体囊外摘出术,建立兔晶状体后囊膜混浊(PCO)模型,术毕kistrin注射组囊袋内注入80 mg/L kistrin 0.2 ml,生理盐水对照组注入等量生理盐水.术后1、3、5、7、14 d,在裂隙灯下观察实验动物晶状体PCO情况,并按照Odrich法进行分级.术后14d及3个月分别处死两组兔各6只,取出晶状体进行常规石蜡切片,行苏木精-伊红染色,光学显微镜下观察晶状体病理改变;行Masson染色观察晶状体囊袋内胶原纤维增生情况,免疫组织化学法检测兔晶状体后囊Ⅳ型胶原的表达. 结果 术后14 d,生理盐水对照组与kistrin注射组各级PCO的眼数差异无统计学意义(P=0.093),术后1、2、3个月,生理盐水对照组形成2～3级PCO的眼数明显多于kistrin注射组,差异均有统计学意义(P=0.041、0.014、0.022).晶状体组织学检查表明,术后14 d生理盐水对照组LECs层数明显多于kistrin注射组,瞳孔区后囊膜可见单层细胞黏附,kistrin注射组后囊则保持光滑,术后3个月可见生理盐水对照组晶状体后囊的LECs转化为纤维细胞,kistrin注射组较少见.Masson染色显示术后3个月生理盐水对照组晶状体前囊膜撕囊口处与后囊膜之间胶原纤维的蓝绿色染色明显多于kistrin注射组.免疫组织化学染色表明,术后14 d及30 d,生理盐水对照组晶状体后囊膜Ⅳ型胶原的灰度值均明显低于kistrin注射组,差异均有统计学意义(P=0.000、0.001). 结论 去整合素kistrin能够抑制兔眼晶状体囊外摘出术术后晶状体后囊LECs和Ⅳ型胶原增生.     

容积调控性氯通道与晶状体后囊膜混浊发病机制的关系

后囊膜混浊为白内障摘除术后残留在晶状体囊袋赤道部的晶状体上皮细胞异常增生移行至后囊膜并分化为成纤维细胞所致,其可直接影响患者术后视力及视觉质量.最近研究发现晶状体上皮细胞上表达多种类型的氯通道,其中容积调控性氯通道作为生物体内一类重要的阴离子通道,除了与细胞的容积调节有关,还在细胞增生、移行与分化等过程中起着重要作用,与晶状体后囊膜混浊形成相关.深入了解此通道可为术后晶状体后囊膜混浊的治疗和预防提供新的线索.     

Nd：YAG激光治疗白内障术后囊袋阻滞综合征疗效分析

目的:探讨白内障超声乳化吸出联合人工晶状体植入术后晶状体囊袋阻滞综合征临床特征及Nd:YAG激光治疗方法选择.方法:回顾性分析7例术后晶状体囊袋阻滞综合征临床特征及Nd:YAG激光治疗方法.其中2例术后早期晶状体囊袋阻滞综合征采用Nd:YAG激光前囊膜周边造孔术;4例术后晚期晶状体囊袋阻滞综合征仅晶状体后囊膜混浊、增厚,采用Nd:YAG激光后囊膜切除术;1例术后晚期晶状体囊袋阻滞综合征伴有人工晶状体前、后表面光学部纤维增殖膜,采用Nd:YAG激光人工晶状体前后囊膜切除术后再行晶状体后囊膜切除术.结果:患者7例经Nd:YAG激光治疗后均恢复较好视力,未见严重并发症.结论:根据术后囊袋阻滞综合征临床特征采取相应的Nd:YAG激光治疗安全有效.     

A new model of posterior capsule opacification in rodents

PURPOSE: To describe a new model of posterior capsule opacification (PCO) in rodents METHODS: An extracapsular lens extraction (ECLE), by continuous curvilinear capsulorrhexis and hydrodissection, was performed in 42 consecutive Brown Norway rats. Animals were killed at 0, 6, and 24 hours and 3, 7, and 14 days after surgery. Eyes were enucleated and processed for light microscopy and immunohistochemistry. RESULTS: In 34 (81%) of the animals the operated eye appeared well healed before death, with a clear cornea and a well-formed anterior chamber. In eight (19%) there was no view of anterior segment structures because of hyphema, fibrin, or corneal opacification. PCO was clinically evident 3 days after ECLE and was present in all animals at 2 weeks. Immediately after ECLE, lens epithelial cells (LECs) were present in the inner surface of the anterior capsule and lens bow. Twenty-four hours after surgery, LECs started to migrate toward the center of the posterior capsule. At 3 days, multilayered LECs, some spindle shaped, were present throughout the lens capsule. Capsular wrinkling was apparent. Lens fibers and Soemmering's ring were observed in all animals 14 days after surgery, indicating some degree of cellular differentiation. Activated macrophages were present in greater numbers at 3 and 14 days after surgery (P < 0.05), when proliferation and migration of LECs appeared to be greatest, and lens fiber differentiation was evident, respectively. CONCLUSIONS: In rodents PCO occurs after ECLE and is associated with low-grade inflammation, mostly of mononuclear macrophages. Although no intraocular lens implantation was performed, this model appears to be valuable for studying the sequence of events that leads to PCO after cataract surgery and the extracellular matrix cues that promote lens fiber differentiation.     

直角边缘人工晶状体预防兔后囊膜混浊的实验研究

目的探讨直角边缘人工晶状体（intraocular lens，IOL）预防后囊膜混浊（posterior capsule opacification，PCO）的作用。方法30只新西兰兔进行超声乳化晶状体摘出联合囊袋内IOL植入术后，随机植入Crane OV-55CP、Crane OV-55C、Alcon TYPE 5C 3种IOL之一。观察术后并发症和PCO情况。术后3月行光镜和透射电镜检查，观察晶状体后囊膜的形态学变化。结果术后3月Crane OV-55CP组的PCO程度比Crane OV-55C和Alcon TYPE 5C组轻（P〈0．05），各组Soemmefing环形成程度无差异（P〉0．05）。病理学检查发现Crane OV-55CP组兔赤道部增生的晶状体上皮细胞在人工晶状体的直角边缘处受到了阻挡。Crane OV-55C和Alcon TYPE 5C组大量晶状体上皮细胞迁移至后囊膜。结论直角边缘IOL延缓了兔PCO的发生、发展，是预防PCO简便安全有效的方法。     

Lens epithelial cell regression on the posterior capsule with different intraocular lens materials

BACKGROUND/AIMS—Posterior capsular opacification (PCO) is caused by proliferation and migration of lens epithelial cells (LECs) across the posterior capsule and is the commonest cause of reduced vision after cataract surgery. The influence of intraocular lens (IOL) material on the process of LEC migration was studied.
METHODS—90 eyes underwent standardised extracapsular surgery, with capsulorhexis and "in the bag" IOL placement. They were randomised to receive a three piece 6 mm lens of PMMA, silicone, or polyacrylic (AcrySof, Alcon, Fort Worth, TX, USA). On days 7, 30, 90, 180, and years 1 and 2 high resolution digitised retroillumination images were taken of the posterior capsule. The presence of LECs was determined at 90 days and 2 years, and their progression or regression was established by serial examination of images.
RESULTS—LECs were seen in 93% of silicone and 97% of PMMA IOLs at 90 days, compared with 46% of polyacrylic (p<0.001). At year 2 LECs were present in all patients with silicone or PMMA lenses, whereas 62% of patients with polyacrylic IOLs had LECs (p<0.001). Of those patients with LECs at day 90 LEC regression occurred in 8% with silicone IOLs and 15% of PMMA cases, compared with 83% of patients with polyacrylic IOLs (p<0.0001).
CONCLUSION—The presence of LECs on the posterior capsule was considerably lower with polyacrylic than PMMA or silicone IOLs and LEC regression occurred more frequently. The lower incidence of LECs and the higher rate of regression may explain why PCO formation appears to be reduced with polyacrylic lenses. This has important clinical implications for the prevention of PCO.

Keywords: lens; cell; implant; posterior capsular opacification     

Use of a polylysine-saporin conjugate to prevent posterior capsule opacification.

PURPOSE: To determine the feasibility of applying a polylysine-saporin (PLS) conjugate to the lens capsule at surgery to prevent lens epithelial cell (LEC) proliferation and posterior capsule opacification (PCO). SETTING: Department of Research & Development, Bausch & Lomb Surgical, and Department of Ophthalmology, Saint Louis University, St. Louis, Missouri, USA. METHODS: Fluorescein-labeled polylysine was applied to the lens capsule of rabbits after phacoemulsification and analyzed histologically to determine the extent of binding to the lens capsule and surrounding tissues. The cytotoxin saporin was conjugated to polylysine using bifunctional cross-linkers. This PLS conjugate was applied to LECs in culture and to the lens capsules of rabbits. These eyes were monitored for PCO. RESULTS: Polylysine primarily bound to the lens capsule membranes, with little or no binding to surrounding tissues. When PLS was added to LECs in culture, it was internalized and destroyed the cells. Of 9 rabbit eyes treated with PLS during surgery, 1 remained free of PCO for the life of the animal (40 weeks), while 6 showed a delay of cortical regrowth approximately 2 to 3 times that of control eyes. CONCLUSIONS: Polylysine bound selectively to the lens capsule membrane. The PLS conjugation resulted in a toxic agent that targeted the lens capsule and destroyed proliferating LECs. The application of a PLS conjugate during surgery may prevent PCO.     

Effect of TGF-beta2 and anti-TGF-beta2 antibody in a new in vivo rodent model of posterior capsule opacification

PURPOSE: This study evaluated the effect of transforming growth factor (TGF)-beta2 and anti-TGF-beta2 antibody in a rodent model of posterior capsule opacification (PCO). METHODS: An extracapsular lens extraction (ECLE) was performed in 72 Sprague-Dawley rats. At the end of the procedure, 10 microL TGF-beta2 (TGF-beta2-treated group), fetal calf serum (FCS)/phosphate-buffered saline (PBS; FCS/PBS-treated control group), a human monoclonal TGF-beta2 antibody (anti-TGF-beta2-treated group), or a null control IgG4 antibody (null antibody-treated control group) was injected into the capsule. Animals were killed 3 and 14 days postoperatively. Eyes were evaluated clinically prior to euthanatization, then enucleated and processed for light microscopy and immunohistochemistry afterward. PCO was evaluated clinically and histopathologically. Student's t-test and chi(2) were used to assess differences between groups. RESULTS: There were no statistically significant clinical or histopathological differences in degree of PCO between the TGF-beta2- and FCS/PBS-treated groups at 3 and 14 days after ECLE. Nor were there differences between the anti-TGF-beta2- and the null antibody-treated groups, with the exception of the histopathology score for capsule wrinkling 3 days after ECLE (P = 0.02). alpha-Smooth-muscle actin staining was observed in the lens capsular bag only in areas where there was close contact with the iris. CONCLUSIONS: No sustained effect of TGF-beta2 or anti-TGF-beta2 antibody on PCO was found in rodents at the dose and timing administered in this study. Iris cells may play a role in the process of epithelial mesenchymal transition linked to PCO.     

骆驼蓬总碱及其脂质体对兔晶状体上皮细胞的抑制作用

探讨骆驼蓬总碱及其脂质体对兔后囊混浊形成的影响。方法 用改良冻融法制备ＬＴＡＨ。１０只健康纯种新西兰白兔分别每眼行晶状体囊外摘出术并分别注入０．１ｍｌ浓度为０．２ ＴＡＨ及ＬＴＡＨ。观察术后后囊，手术前后眼压，角膜内皮细胞，视网膜电图及组织病理学变化。