乙二胺四乙酸二钠对晶状体上皮细胞清除作用的实验研究

目的:探讨乙二胺四乙酸二钠(EDTA)对晶状体囊膜上皮细胞的清除效果。方法:收集白内障超声乳化囊外摘除术中环形撕取的晶状体前囊膜32例,将其随机分为4组,分别用5mmol/L,10mmol/L,20mmol/L EDTA及生理盐水进行前囊膜消化处理,观察晶状体上皮细胞的清除情况。结果:经过20mmol/L EDTA溶液处理的晶状体前囊膜,上皮细胞可以完全脱落,经过5mmol/L,10mmol/L EDTA溶液处理的晶状体前囊膜,上皮细胞部分脱落,生理盐水对照组仅见少量晶状体上皮细胞脱落。结论:EDTA可以清除晶状体囊膜的上皮细胞,并随浓度增加效果增强,该结果可为临床应用EDTA清除晶状体上皮细胞以防治后发性白内障(posterior capsular opacification,PCO)的发生提供实验基础和理论依据。     

依地酸二钠预防后发性白内障的实验研究

目的 探讨应用依地酸二钠(EDTA)清除晶状体上皮细胞,预防后发性白内障的可行性.方法 白内障摘除术中连续环形撕囊后取得人晶状体前囊膜30例,随机分为五组,分别用平衡盐溶液(对照组、A组),5mmol/L、10 mmol/L、15 mmol/L、30 mmol/L的EDTA溶液(B、C、D、E组)进行处理,观察晶状体上皮细胞的清除情况.结果 A组未见明显细胞脱落;B组晶状体上皮细胞仅有少部分脱落;C组和D组晶状体上皮细胞大部分脱落;E组晶状体上皮细胞完全脱落.结论 合适浓度的EDTA可以作为白内障手术中清除晶状体上皮细胞的有效方法,为防治后发性白内障提供理论依据.     

白内障术后后囊浑浊发生机制的研究--老年性白内障晶状体上皮细胞的体外培养

目的观察老年性白内障晶状体上皮细胞体外培养的生长特点。为研究老年性白内障及术后后囊浑浊的发生机制及防治奠定基础。方法应用改良组织块培养法对超声乳化术中老年性白内障晶状体前囊上皮细胞进行体外培养，在倒置显微镜下观察其生长和分化的规律。结果前囊接种3—5天，有新生上皮细胞自囊片的边缘长出并向四周延伸，第3-4周部分细胞内出现空泡和颗粒等结构改变，生长近于停止；传代培养细胞不能增生。结论老年性白内障晶状体前囊上皮细胞体外增生能力有限。改良组织块培养法培养晶状体上皮细胞简单有效。     

老年性白内障与透明晶状体上皮内蛋白酶体活性的比较

目的:比较老年性白内障晶状体上皮与透明晶状体上皮内蛋白酶体的糜蛋白酶样、胰蛋白酶样酶活性是否存在差异,进而探讨蛋白酶体在老年性白内障发生发展中的作用机制。方法:白内障晶状体囊膜取自在我院行白内障超声乳化及人工晶状体植入的患者,透明晶状体囊膜取自广东省眼库,年龄与白内障病人相匹配。反复冻融囊膜使细胞脱落,将囊膜从溶液中除去。各组样品中分别加入LLVY或VGR,在不同时间测定荧光强度。结果:透明晶状体上皮的蛋白酶体活性明显高于白内障组(P<0.01);皮质型与非皮质型白内障晶状体上皮的糜蛋白酶样酶活性无明显区别(P>0.05);MG-132明显抑制各组蛋白酶体的活性。结论:与透明晶状体相比,白内障晶状体上皮内蛋白酶体的糜蛋白酶样、胰蛋白酶样酶活性明显降低,蛋白酶体活性降低可能在老年性白内障的形成中起重要的调节作用。     

骨桥蛋白在白内障晶状体上皮细胞中的表达

目的：探讨骨桥蛋白（osteopontin，OPN）在白内障晶状体上皮细胞中的表达水平及作用。方法：采用免疫组织化学法检测22例前囊下性白内障、17例核性白内障、14例皮质性白内障患者和11例正常晶状体上皮细胞中OPN的表达，并进行阳性细胞计数和各组间比较。结果：OPN在前囊下白内障患者晶状体上皮细胞中的表达明显增多，其阳性表达率与核性白内障、皮质性白内障及正常人晶状体比较，差异有显著意义（P〈0．01）。结论：OPN在前囊下性白内障晶状体上皮细胞中表达阳性，在前囊下白内障的形成过程中可能起重要作用。     

晶状体前囊染色在白内障手术中的应用

目的 探讨晶状体前囊染色在白内障手术中应用。方法 在６５２眼白内障摘出术中,采用０．５％吲哚青绿溶液进行晶状体前囊染色后,实施连续环形撕囊术或截囊术。结果 所有病例在吲哚青绿溶液注入前房后,晶状体前囊染成浅绿色,在手术显微镜下清晰可辨,使撕囊或截囊术得以快速顺利完成,并且不干扰其他手术步骤进行,术后无不良反应。结论 吲哚青绿溶液晶状体囊染色有助于白内障手术中辨认晶状体前囊。     

年龄相关性白内障晶状体上皮细胞的超微结构研究

目的:研究年龄相关性白内障晶状体上皮的超微结构变化。方法:用日本产HU-12A型电镜对8例年龄相关性白内障及3例正常透明晶状体上皮细胞的超微结构进行观察。结果:7例年龄相关性白内障晶状体前囊上皮细胞形状不规则,相邻细胞间以许多复杂的指状突超紧密镶嵌,部分上皮细胞内水肿,细胞间隙亦水肿呈较大的空泡状,细胞内线粒体等细胞器肿胀,个别区域上皮细胞坏死脱落,1例年龄相关性白内障晶状体前囊上皮细胞体积固缩,细胞核染色质分裂并出现凝聚的凋亡小体。结论:晶状体上皮细胞超微结构的病理形态学改变是年龄相关性白内障发生发展的重要原因之一。     

波形纤维蛋白在老年性白内障晶状体上皮细胞的表达

目的观察波形纤维蛋白在老年性白内障晶状体上皮细胞的变化.方法用卵白素-生物素过氧化物酶法对22例老年性白内障患者晶状体前囊膜上皮细胞进行波形纤维蛋白染色,采用包埋前免疫酶电镜技术处理6个晶状体前囊膜标本,并观察其超微结构;利用十二烷基硫酸钠聚丙烯酰胺凝胶电泳及Westemnblot法分析4个晶状体表层组织(前囊膜、上皮细胞和表层皮质)中的波形纤维蛋白.结果老年性白内障晶状体上皮细胞的波形纤维蛋白表达减弱,与对照组比较差异有显著性(t=2.0948,P<     

老年性和糖尿病性白内障晶状体上皮细胞的超微结构改变

目的 观察正常人，老年性白内障、糖尿病性白内障晶状体上皮细胞的超微结构，探讨细胞凋亡在白内障发生中的作用。方法 收集正常人、老年性白内障、糖尿病性白内障晶状体前囊膜送透射电镜扫描，取老年性白内障和糖尿病性白内障前囊膜各15例，用脱氧核苷酸末端转移酶缺口标记原位细胞检测法，检测晶状体上皮细胞的凋亡细胞。结果 老年性和糖尿病性白内障晶状体上细胞的超微结构在形态上为扁平状，胞浆出现空泡变性，胞核出现固缩，染色质边聚，浓缩等改变。糖尿病性白内障晶状体上皮细胞的胞浆空泡变性更加严重。在光镜下老年性和糖尿病性白内障的晶状体上皮均有凋亡细胞。结论 老年性白内障和糖尿病性白内障的发生与晶状体上皮的细胞凋亡密切相关。     

水通道蛋白1在老年性白内障及透明晶状体前囊膜上皮细胞的表达

目的明确水通道蛋白1(aquaporin1,AQP1)在透明晶状体及老年性白内障晶状体上皮细胞及前囊膜的表达及分布,并初步探讨AQP1在老年性白内障发病中的重要作用。方法收集52例老年性白内障及10例正常透明晶状体前囊膜,通过HE染色和免疫组化方法检测AQP1的表达。结果老年性白内障及正常透明晶状体上皮细胞中均可见AQP1的表达,透明晶状体组中AQP1的阳性细胞表达率(100%)显著高于老年性白内障组(53.8%)(P<0.05)。而在晶状体纤维中未见表达。结论晶状体上皮细胞是晶状体代谢最活跃的部位,终身进行有丝分裂,并移行至赤道部分化成晶状体纤维。晶状体上皮细胞有AQP1的活性表达,这对于维持晶状体上皮细胞一定的增殖潜能,保证晶状体的生理动态平衡和晶状体的透明性起到了重要的作用。提示AQP1在老年性白内障晶状体前囊膜及上皮细胞中的异常表达与老年性白内障的发生密切相关,具体调控机制有待进一步研究。     

硅油眼晶状体前囊膜及上皮细胞的病理学改变及Caspase-3表达

目的 观察硅油充填眼晶状体前囊膜及上皮细胞的病理学改变及Caspase—3表达，探讨硅油引起白内障的病因及与凋亡的关系。方法 收集27只晶状体前囊膜，通过HE染色和免疫组化方法对硅油眼的晶状体前囊膜和上皮细胞及Caspase—3在其上的表达进行观察。结果 前囊膜下许多空间被硅油滴占据；Caspase—3在晶状体上皮细胞有很好的表达，27例中有23例呈阳性表达。结论 硅油滴与白内障的发生、发展有关；凋亡在硅油眼并发白内障中存在。     

老年性白内障晶状体上皮细胞凋亡及相关基因蛋白的表达

目的 探讨老年性白内障与晶状体上皮细胞凋亡的关系。方法透射电镜下观察老年性白内障晶状体上皮细胞的超微结构；Tunel法检测凋亡细胞百分率；并对其晶状体上皮细胞DNA进行琼脂糖凝胶电泳；免疫组化法检测P53、bcl-2在老年性白内障晶状体上皮细胞中的蛋白表达。结果 透射电镜下发现老年性白内障晶状体上皮细胞中有凋亡细胞；凋亡细胞百分率为8．4％～37．8％，琼脂糖凝胶电泳出现梯状条带；P53在老年白内障晶状体上皮细胞中蛋白表达率为16．9％～19．1％，bcl-2无蛋白表达。结论 老年性白内障的发生可能与其晶状体上皮细胞凋亡有关。     

核因子κB在年龄相关性白内障晶状体前囊膜的表达

目的研究核因子κB（NF-κB）在正常人、年龄相关性白内障患者晶状体前囊膜上皮细胞中表达的差异,探讨NF-κB在年龄相关性白内障发病机制中的作用。方法收集白内障术中60例60眼年龄相关性白内障的前囊膜组织（白内障组）,选取正常供体5例10眼的透明晶状体前囊膜组织作为正常对照组。取白内障组30眼标本和正常对照组5眼标本用于免疫组织化学法检测NF-κBp65蛋白在晶状体前囊膜上皮细胞中的表达;另外白内障组30例标本及正常对照组的5例标本采用实时荧光定量聚合酶链反应（PCR）方法检测NF-κBp65mRNA在晶状体前囊膜上皮细胞中的表达。结果正常晶状体前囊膜上皮细胞细胞质和细胞核中可见NF-κBp65蛋白的弱阳性表达,以细胞质为主;年龄相关性白内障晶状体前囊膜上皮细胞的细胞质和细胞核中NF-κBp65蛋白呈强阳性表达。白内障组NF-κBp65吸光度（A）值为0.1658±0.022,正常对照组为0.2889±0.043,差异有统计学意义（t=6.2109,P〈0.05）。白内障组NF-κBp65mRNA表达量为1.454±0.081,正常对照组为0.951±0.075,差异有统计学意义（t=12.9683,P〈0.05）。结论 NF-κB表达水平的升高与年龄相关性白内障的发病密切相关,其表达异常可能参与年龄相关性白内障的发生发展。     

年龄相关性白内障基因异常表达的研究

目的比较年龄相关性白内障晶状体上皮细胞(LEC)基因表达与正常人的差异,探讨白内障发病的分子机制。方法术中撕取晶状体前囊36眼,其中单纯核性年龄相关性白内障10眼,成熟期年龄相关性白内障19眼,角膜移植供体眼及外伤性晶状体脱位7眼(正常对照)。3组晶状体囊膜分别提取总RNA,将合格的各组RNA样品做差异显示(DD-RT-PCR),寻找差异表达基因,再将其分离、扩增、测序,最后选取与白内障形成密切相关的胰岛素样生长因子结合蛋白(IGFBP-5)及溶酶体相关膜蛋白2(LAMP-2)基因进行验证。结果各组RNA纯度达到要求,总RNA完整,经差异显示,年龄相关性白内障组比正常人组低表达的条带有5条,高表达条带15条。对6条明亮条带测序后,其中3个片段分别与IGFBP-5、LAMP-2及小核糖体蛋白10(rps10)有很高的同源性,其他3个片段均为新近克隆未知功能的基因或基因组序列。验证表明,年龄相关性白内障IGFBP-5表达水平低于正常对照(P<0.01),而年龄相关性核性白内障LAMP-2表达水平高于正常对照。结论IGFBP-5和LAMP-2是年龄相关性白内障相关基因,差异显示技术在研究年龄相关性白内障发生中有重要意义。     

Human and porcine anterior lens capsule as support for growing and grafting retinal pigment epithelium and iris pigment epithelium

· Purpose: To establish a method for transplantation of cultured monolayers of RPE and IPE into the subretinal space, anterior lens capsule was evaluated for its suitability to serve as growth support and carrier for transplantation procedures. · Materials and methods: Twenty-four anterior lens capsules were obtained from porcine eyes. The same number of human lens capsules was obtained during cataract surgery. Six lens capsules of each species were stored at –80°C. Subsequently, the capsules were transferred onto type-I collagen. A second set of six lens capsules was treated identically except for the cryo treatment. A third set of six capsules was initially exposed to 0.05% trypsin for 30 min. Suspended porcine RPE and IPE cells (5×10⁴ cells/well) were seeded on the top of each capsule. The remaining six lens capsules served as controls and were incubated in uncoated 12-well dishes without undergoing experimental treatment. The cultures were maintained in a water-saturated atmosphere at 37°C with 5% CO₂. Six days later, viability, morphology, and cell density were determined. The capsules covered by a confluent monolayer of cells were transferred into uncoated wells and cultivated for another 10 days. At the end of the experiment, light and phase-contrast microscopy was performed on all capsules. · Results: Storage at –80°C and exposure to trypsin resulted in significant reduction of cellular contamination. The highest cell density was found after 5 days when capsules which had undergone cryopreservation or trypsin exposure served as support for RPE and IPE. The pigment cell layer was firmly attached to the capsules and permitted a transfer to other culture flasks without significant cell loss. The IPE cell layer remained confluent after transfer to uncoated culture flasks, while the RPE cell layer ceased to proliferate 10 days after transfer. · Conclusions: Lens capsules may be suitable for growing and supporting monolayers of pigment epithelial cells. Especially IPE cells formed stable monolayers on anterior lens capsules which could be transferred to secondary culture flasks without inflicting damage on the cells. Received: 6 October 1998 Revised version received: 18 December 1998 Accepted: 30 March 1999     

老年性和糖尿病性白内障晶状体上皮细胞的超微结构改变

目的 观察正常人,老年性白内障、糖尿病性白内障晶状体上皮细胞的超微结构,探讨细胞凋亡在白内障发生中的作用。方法 收集正常人、老年性白内障、糖尿病性白内障晶状体前囊膜送透射电镜扫描,取老年性白内障和糖尿病性白内障前囊膜各15例,用脱氧核苷酸末端转移酶缺口标记原位细胞检测法,检测晶状体上皮的凋亡细胞。结果 老年性和糖尿病性白内障晶状体上皮细胞的超微结构在形态上为扁平状,胞浆出现空泡变性,胞核出现固缩,染色质边聚,浓缩等改变。糖尿病性白内障晶状体上皮细胞的胞浆空泡变性更加严重。在光镜下老年性和糖尿病性白内障的晶状体上皮均有凋亡细胞。结论 老年性白内障和糖尿病性白内障的发生与晶状体上皮的细胞凋亡密切相关。     

The human anterior lens capsule--an attempted chemical debridement of epithelial cells by ethylenediaminetetracetic acid (EDTA) and trypsin.

The addition of edetic acid (EDTA) or trypsin to the infusion during a simulated extracapsular cataract extraction on cadaver eyes facilitates the removal of lens epithelial cells from the anterior capsule. Modification of the chemical composition of infusions used during extracapsular surgery may maximise lens epithelial cell removal and hence reduce the incidence of opacification of the posterior capsule after cataract extraction.     

Specular microscopy of the anterior lens capsule after endocapsular lens implantation

Endocapsular phacoemulsification and complete in-the-bag intraocular lens (IOL) fixation was performed on 77 eyes. Specular microscopy of the anterior lens capsule was performed postoperatively, and the process of anterior capsule opacification after endocapsular cataract surgery was classified. Lens epithelial cells on the inner surface of the anterior lens capsule, facing the optical portion of the IOL, underwent a diffuse fibrosis. In more peripheral areas, where the anterior and posterior capsules were tightly adherent, the regularly proliferating lens epithelial cells retained their polygonal shape. These findings suggest the possibility of retaining the transparency of the anterior lens capsule postoperatively by refilling the capsular bag to apply internal pressure.     

Lens epithelial cell death secondary to acanthamoeba keratitis: absence of capsular bag opacification six years after cataract surgery

Purpose

To show the evolution of anterior chamber structures 6 years after cataract surgery in a case with Acanthamoeba keratitis (AK).

Methods

A 37-year-old woman with AK receiving long-term treatment with chlorhexidine, propamidine isethionate and steroids developed a white cataract and iris atrophy. Penetrating keratoplasty and cataract surgery were performed with subsequent intraocular pressure elevation requiring Molteno shunt implantation. Two years after the last surgery, endothelial decompensation developed and another penetrating keratoplasty was performed. Intraoperatively, the anterior and posterior capsules were completely transparent.

Results

Six years after cataract surgery, the intraocular lens was centered with clear anterior and posterior capsules without lens epithelial cells proliferation. No Soemmering''s ring formation or posterior capsule opacification was found. Also, no zonular damage or pseudophacodonesis was observed.

Conclusions

This case suggests that AK infection and AK treatment not only cause white progressive cataract but also lens epithelial cell death. The capsules may be completely clear 6 years after cataract surgery, with a good quality of vision regardless of intraocular lens material or design.Key words: Acanthamoeba keratitis, Lens epithelial cells, Penetrating keratoplasty, Glaucoma, Cataract, Iris atrophy     

Results of 87 cases of intraocular lens implantation at the front of the whole capsular bag

To prevent or delay Elschnig pearl formation, we performed an unusual surgical technique on 87 patients: intraocular lens implantation at the front of the whole capsular bag. We tried to obtain tight adhesion between the anterior and posterior capsules so the lens epithelial cells could not proliferate and form Elschnig pearls. In fact, the patients exhibited no Elschnig pearls or Soemmering's ring, but developed large anterior capsular folds and a multilayer cell membrane between both capsules after two (three cases), six (19 cases), and 12 months (14 cases). This led to a decrease in visual acuity, and a Nd:YAG laser anterior and posterior capsulotomy was required in most cases (83%). Several months later, these eyes developed enormous Elschnig pearls emanating centrifugally from the laser opening, strongly suggesting that lens epithelial cells require aqueous humor for Elschnig pearl transformation. Since it is not yet possible to obtain perfect adhesion between anterior and posterior capsules, our results confirm the need to remove the anterior capsule after extracapsular cataract extraction.