人工晶状体袢的角度与后发性白内障相关性研究

目的　探讨后发性白内障与人工晶状体袢角度之间的关系。方法　 9只科研兔随机分为 3组 ,每组 6眼。麻醉后 ,行透明晶状体囊外摘出术 ,并分别植入袢角度不同的PMMA人工晶状体 ,第 1组植入 0°袢人工晶状体 ,第 2组植入 5°袢人工晶状体 ,第 3组植入 10°袢人工晶状体。术后 6个月 ,裂隙灯下观察晶状体后囊混浊情况并分级 ,取出晶状体后囊 ,行光镜及透射电镜检查 ,并采用免疫组化法对晶状体后囊染色 ,然后采用医用多功能图像分析软件对晶状体后囊表面及赤道部的增殖细胞核抗原 (proliferatingcellnucle arantigen ,PCNA)进行定量检测 ,用SPSS统计软件分析结果。结果　裂隙灯下可见术后随着人工晶状体袢角度的增加 ,发生后发性白内障的例数及等级均呈下降趋势。光镜及透射电镜扫描可见发生后发性白内障的晶状体后囊表面有一层或多层晶状体上皮细胞生长 ,未发生后发性白内障的晶状体后囊表面无晶状体上皮细胞生长 ,赤道部均可见多层晶状体上皮细胞生长。PCNA定量结果 :植入 0°袢人工晶状体组与植入 5°袢人工晶状体组之间后囊表面及赤道部PCNA阳性率 (分别为 10 .5 0 0±2 2 5 8,9.16 7± 2 .2 2 9)无明显差异 (P >0 .0 5 ) ,而植入 10°袢人工晶状体组 (4.5 0 0± 1.871)与植入 0°袢人工晶状体组及植入     

细胞浆信息传递介质Smad 4在兔晶状体上皮细胞增殖中的作用

目的：探讨细胞浆信息传递介质Smad4在兔晶状体上皮细胞增殖及转化中的作用。方法：将健康白色家兔63只随机分成实验组（49只）及对照组（14只）,实验组兔双眼行透明晶状体皮质吸除术,对照组兔不做处理。在术后1及3d,1周、1、2、3及5个月分别处死7只实验组兔及2只对照组兔,分别应用原位杂交及免疫组化技术检测晶状体上皮细胞中Smad 4mRNA及增殖细胞核抗原（proliferating cell nuclear antigen,PCNA）的表达。结果：正常兔晶状体上皮细胞胞浆中Smad 4mRNA表达呈弱阳性。术后1d晶状体赤道部及前囊下部Smad 4mRNA无表达,吸光度（A）值与术前相比,差异有显著意义（P＜0．01）;术后3d Smad 4mRNA呈阳性表达,且随时间延长表达逐渐增强,晶状体赤道部细胞在术后1周时Smad 4mRNA表达最强;晶状体前囊下部细胞在术中1个月时Smad 4mRNA表达最强。PCNA的表达在术后1d最强,之后逐渐减弱,术后1个月时恢复至术前水平。术后1－2个月,晶状体赤道部出现转化的成纤维细胞,Smad 4mRNA呈阳性表达。结论：Smad 4mRNA在正常晶状体上皮细胞胞浆中有表达,Smad 4参与晶状体上皮细胞的增殖代谢过程,可能促使晶状体上皮细胞转化为成纤维细胞。     

结缔组织生长因子和转化生长因子与后发性白内障发病关系的研究进展

后发性白内障（亦称后囊膜混浊）是白内障术后常见并发症,由术后残留的晶状体上皮细胞在囊膜上转分化、增生、移行,产生胶原所致。结缔组织生长因子（connective
tissue growth factor,
CTGF）在正常晶状体上皮细胞中不表达,而在前囊膜下型白内障及后囊膜混浊时大量表达,产生胶原,参与前囊膜下型白内障和后发性白内障的发生、发展过程。
TGF（transforming growth
factor,TGF） β诱导晶状体上皮细胞转分化、移行、过度增殖,是导致前囊膜下型白内障和后发性白内障发生的重要因素。CTGF与TGF β主要通过Smad信号通路诱导晶状体上皮细胞发生转分化。CTGF是TGF β的下游效应因子,TGF β调控CTGF的表达。     

后发性白内障的发病机制及其防治研究进展

晶体后囊混浊是白内障囊外摘除术后的主要并发症之一,它直接影响白内障术后视力的恢复。自Ｖａｎ等１９５９年培养成功动物晶体上皮细胞、Ｈａｍａｄａ等在１９７８年培养成功人晶体上皮细胞以来国内外学者做了大量的研究,从细胞水平探讨晶体后囊混浊的发生机制,治疗及预防方法。 一、后发性白内障的组织病理学研究 正常人晶体上皮细胞仅限于前囊下,赤道部及赤道弓部,为单层立方上皮,分为三个不同的生物活性区域;中央区、赤道前区和赤道区。有丝分裂在年轻人的晶体分布于整个前囊,而在成人,大多数有丝分裂发生于赤道前区。晶体上皮…     

bcl-2基因和增殖细胞核抗原在人晶状体上皮细胞中的表达

目的：观察正常胚胎、儿童和老年人晶体上皮细胞中bcl-2基因和增殖细胞核抗原（proliferating cell nuchlear antigen,PCNA)的表达状况,探讨其与晶状体上皮细胞增殖潜能的关系。方法采用免疫组织化学染色法光谱下检测胚胎组（胎龄5－10个月）、儿童组（7个月至13岁）及老年组（＞50岁）晶状体上皮细胞中bcl-2基因和PCNA的表达状况。结果：bcl-2基因和PCNA在胚胎组和儿童组晶状体上皮细胞中表达的阳性率显著高于老年组（P＜0．01）,且以晶状体赤道部明显,结论日 状体上皮细胞的增殖潜能以及后发性白内障的发生与晶状体上皮细胞内bcl-2基因及PCNA的表达有关。     

去整合素抑制兔晶状体上皮细胞增殖的实验研究

目的 探讨去整合素(Kistrin)对兔眼后发性白内障发生过程中晶状体上皮细胞(lens epithelial cells,LEC)增殖的抑制作用.方法 雄性新西兰大白兔36只,随机分为对照组:A组,实验组:B、C、D组,每组各9只兔9眼(右眼).4组均行透明晶状体囊外摘出术,术毕A组兔眼前房注入林格液0.2 mL,B、C、D组分别注入40 μg·L-1、80μg·L-1和160 μg·L-1 Kistrin溶液0.2 mL.术后第14天采用免疫组织化学法对4组兔后发性白内障发生过程中LEC增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)的表达进行检测,计算增殖指数(proliferative index,P1)值,PI=PCNA阳性细胞数/50×100%;通过HE染色及透射电镜观察角膜改变情况.结果 (1)术后第14天PI值A组:0.320±0.020、B组:0.175±0.030、C组:0.065±0.020、D组:0.190±0.050,B、C、D组较A组PI值均明显降低(均为P＜0.01);C组较B、D组PI值均明显降低(均为P＜0.01),B组与D组PI值比较差异无统计学意义(P＞0.05);(2)术后第14天赤道部PI值A组:0.390±0.060、B组:0.230±0.040、C组:0.110±0.030、D组:0.240±0.080;中央部PI值A组:0.250±0.050、B组:0.120±0.030、C组:0.060±0.020、D组:0.140±0.040,4组赤道部较中央部PCNA阳性表达均明显升高(均为P＜0.01).(3)实验组C组与A组比较:角膜内皮细胞超微结构无明显改变.结论 40～160μg·L-1的Kistrin灌注液可有效地抑制制兔眼后发性白内障的发生;赤道部LEC增殖能力强于中央部;80μg·L-1可作为进一步研究Kistrin抑制后发性白内障形成的较适宜浓度.     

PCNA在不同年龄组人晶状体上皮细胞中的表达

目的　检测不同年龄组人晶状体上皮细胞增殖细胞核抗原 (PCNA)表达 ,探讨白内障术后残留晶状体上皮细胞的增殖与后囊混浊的关系及儿童后发性白内障高发的原因。方法　按年龄段分为小儿组 (<12岁 )及老年组 (5 1～80岁 )。在白内障手术时环形撕囊后获取晶状体中央区前囊膜 ,用免疫组化染色及真彩色医学图像分析系统检测PCNA的表达及积分光密度值 ,取其均值进行统计学分析。结果　定性检测小儿组 10张切片中 7张阳性 ,2张可疑 ,1张阴性。老年组 2张可疑 ,8张阴性。定量检测 (积分光密度值 )小儿组为 2 89± 0 5 7,老年组为 2 13± 0 63 (P <0 0 5 )。结论　小儿组晶状体上皮细胞PCNA的表达明显高于老年组 ,提示可能为小儿后发性白内障高发的重要因素。通过PCNA表达的检测可能为判断后囊混浊高危眼提供依据     

CD44在人晶状体上皮细胞中的表达

目的 研究人晶体状上皮细胞CD44的表达，探索后发障的成因。方法 在白内障超生乳化术中环形撕囊后获得晶状体前囊，立即进行免疫组织化学染色，光镜下观察标本。结果 晶状体上皮细胞结构完整，30例中25例染色呈阳性。结论 人晶状体上皮细胞原位表达CD44。CD44可能参与白内障术后残余晶状体上皮细胞的移行、黏附致后囊混浊这一过程，抑制细胞黏附因子CD44也许可以成为治疗后发障的又一新途径。     

后发性白内障

后发性白内障（简称后发障）是现代白内障囊外摘出手术后影响视力最常见的并发症，手术后残留的晶体上皮细胞增殖、生成新的晶体物质或／和纤维化生引起后囊皱缩而混浊。有学者观察了手术后晶体纤维再生的组织学过程，及生化因素对晶体上皮细胞增殖分化的影响，已经发现多种生长因子可影响晶体上皮细胞的有丝分裂活性。进一步研究术后早期晶体上皮细胞的增殖反应机制，对预防后发障的形成有特殊意义。     

大直径环形撕囊近赤道前囊抛光预防后发性白内障

目的 观察白内障超声乳化吸出术中较大直径连续环形撕囊联合近赤道前囊下抛光对预防后发性白内障的作用.方法 对96例(106眼)白内障行超声乳化吸出联合人工晶状体植入术.A组53眼连续环形撕囊直径5.5～6.5mm,常规后囊抛光后用弯注吸针头行近赤道前囊下抛光,植入亲水性丙烯酸酯折叠人工晶状体.B组53眼撕囊直径4.5～5.5mm,不进行前囊抛光,余同A组.观察两组后发性白内障情况.结果 随访2年,术后3个月、6个月两组后发性白内障发生率无明显差异、术后1年A组后发性白内障发生率明显低于B组(P＜0.05).结论 较大直径环形撕囊联合近赤道前囊下抛光,能有效降低后发性白内障的发生.     

碱性成纤维细胞生长因子基因在胎儿与白内障患者晶状体上皮细胞内表达的比较

目的:研究碱性成纤维细胞生长因子基因在胎儿和白内障患者晶状体上皮细胞内表达的区别。方法:采用原位杂交方法,用cDNA探针检测胎儿培养及组织切片中的晶状体上皮细胞和白内障患者前囊中的晶状体上皮细胞的bFGF的mRNA,并用图像分析进行相对定量,比较胎儿培养细胞、组织切片细胞及患者囊膜细胞的积分光吸收度值。结果:胎儿的培养及组织切片中晶状体上皮细胞和白内障患者前囊中的晶状体上皮细胞都存在bFGF基因表达。胎儿体外培养晶状体上皮细胞、胎儿组织切片晶状体上皮细胞和白内障患者晶状体上皮细胞积分光吸收度值分别为627.1±268.7,131.5±42.8和79.2±26.3。胎儿体外培养晶状体上皮细胞积分光吸收度值显著高于胎儿组织切片晶状体上皮细胞(P<0.01);白内障患者晶状体上皮细胞积分光吸收度值显著低于胎儿晶状体上皮细胞(P<0.01)。结论:晶状体上皮细胞体外培养可增加bFGF基因表达;胎儿晶状体上皮细胞bFGF基因表达显著高于白内障患者晶状体上皮细胞。     

Experimental traumatic cataract. II. A transmission electron microscopy and extracellular tracer study

Lens changes caused by injury to the anterior part of the lens were studied with Procion yellow as an extracellular tracer and by transmission electron microscopy at different time intervals after trauma. Both rats and rabbits were used. The findings were related to the slit-lamp appearance of the wounded lenses. In the rat lens a posterior subcapsular cataract developed within the first hour after trauma. Within 1 hr after injury the fluorescent tracer was seen at the wound but was also conspicuous at the posterior pole. Swelling of lens fiber cells and the formation of large syncytical aggregates were found as the posterior opacity enlarged. These changes reached the anterior subcapsular cortex via the equatorial cortex after about 1 month. In the rabbit lens a slight cellular swelling was seen in the subcapsular cortex. Only in one of 15 lenses a posterior subcapsular opacity developed after about 1 week in spite of a large wound. The uptake of Procion yellow was most prominent in the wound area and was never observed at the posterior pole. In both species, no further penetration of the dye occurred through the wound after the epithelium, by regeneration, had sealed the wound. The importance of epithelial wound sealing and that of a restored cellular barrier at the posterior pole are discussed as well as the significance of these factors in the cataract progression.     

人8-羟基鸟嘌呤糖苷酶1与年龄相关性白内障关系的临床研究

目的探讨与年龄相关性白内障（ARC）患者晶状体上皮细胞（LECs）中参与DNA损伤后碱基清除修复途径的氧化损伤修复基因—人8-羟基鸟嘌呤糖苷酶1（HOGG1）水平与ARC的关系。方法收集三种ARC（皮质性、核性、后囊下性）LECs样本,以透明晶状体LECs为对照组,用免疫组化、RT-PCR方法测定HOGG1在LECs的表达情况。结果对照组LECs中可见HOGG1的表达,三种ARC患者LECs中可见HOGG1表达较对照组增高（F=107.62,P〈0.01）,但三种ARC之间没有统计学差异。对照组与ARC组HOGG1均位于细胞质和细胞核。说明ARC LECs的细胞核和细胞质中HOGG1的表达量上调。结论 HOGG1表达上调参与ARC的发生发展。     

EDTA与多聚赖氨酸的铰链物抑制兔后发性白内障的实验研究

目的：探讨EDTA与多聚赖氨酸的铰链物对兔后发性白内障的防治作用。方法：将20只新西兰白兔40眼随机分为A,B,C,D共4组,4组均行透明晶状体囊外摘除术。A组为对照组,术中灌注液为BSS;B,C,D组为治疗组,术中灌注液分别为浓度为10mg/L的EDTA、多聚赖氨酸、EDTA与多聚赖氨酸的铰链物的BSS溶液。2mo后行晶状体后囊膜切片,HE染色统计晶状体上皮细胞（LECs）的密度;并行免疫组织化学染色,用医学图象分析系统检测PCNA表达平均光密度（灰度值OD）。结果：经HE染色,后囊膜LECs密度B和D组较A和C组少,且D组的LECs密度小于B组,均有显著性差异（P〈0.01）;A和C组的LECs密度相差不大,无统计学差异（P〉0.05）。免疫组织化学进行平均光密度测定,A和C组PCNA表达强阳性,B组呈部分阳性表达,D组阳性表达较B组更少。B和D组与A组、B组与D组均有显著性意义（P〈0.01）。A组和C组无显著差异性（P〉0.05）。结论：在活体兔眼中,EDTA、多聚赖氨酸与EDTA的铰链物均有抑制兔LECs增殖的作用,且EDTA与多聚赖氨酸的铰链物组对LECs的抑制作用优于EDTA组,多聚赖氨酸组对LECs的抑制作用不明显。     

Degeneration and transdifferentiation of human lens epithelial cells in nuclear and anterior polar cataracts.

PURPOSE: To study the possible mechanisms of cataractogenesis by evaluating the characteristics of cataractous lens epithelial cells (LECs) in different types of human cataract. SETTING: Kangnam St. Mary's Hospital, The Catholic University of Korea, Seoul, Korea. METHODS: Lens epithelial cells attached to the anterior capsules in eyes with nuclear or anterior subcapsular were analyzed for morphological characteristics by electron microscopy and for cellular characteristics by immunohistochemistry. RESULTS: Human LECs beneath the anterior capsule were degenerated in nuclear cataracts and were transdifferentiated in anterior polar cataracts. In senile nuclear cataractous lenses, LECs beneath the anterior capsule showed degenerative changes in morphology. In nuclear cataracts, LECs were immunohistochemically positive for cytokeratin and vimentin, while those in anterior polar cataracts were positive for vimentin only. The LECs of anterior subcapsular cataracts were transdifferentiated into spindle-shaped fibroblast-like cells without cellular junctions and embedded within a fibrillar meshwork mass. The extracellular matrixes in the anterior capsule of anterior subcapsular cataracts were immunohistochemically positive for fibronectin, laminin, collagen type I, and collagen type IV. CONCLUSIONS: Lens epithelial cells in different types of cataracts have distinct cellular characteristics and may possess a bipotential nature with the ability to transdifferentiate into mesenchymal cells. This may be an underlying mechanism for the development of cataract and capsule opacification.     

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]     

Effects of calcium on lens epithelial cells in rabbits

PURPOSE: The action of lens epithelial cells (LECs) is important for cataract and posterior subcapsular cataract after cataract surgery. In this study, we analyzed the effects of calcium on the characteristics of LECs. METHODS: The LECs were collected using albino rabbits and incubated in minimum essential medium [MEM, Introgen Corp. (12% fetal bovine serum: FBS)] (37 degrees C, 5 % CO2) for a week to induce their proliferation. Cell culture dishes (35 mm) were prepared and 7 mm cylindrical pipes were placed in them. After that, around 10,000 cultured LECs were placed in the pipes and incubated. After 2 hours incubation, the pipes were removed and various doses of MEM (0, 2, 10 and 20 mM) replaced the calcium. Proliferation and shapes of LECs were observed using a confocal microscope and immunohistological analysis [alpha-smooth muscle actin (alpha-SMA) and bromodeoxyuridine (BrdU)]. The LECs were incubated with collagen gel and different calcium doses (0, 2, 10 and 20 mM) of MEM to calculate the contraction rate. RESULTS: It was observed that the LECs changed to fibroblast-like cells at high doses of calcium using a confocal microscope. Histological studies showed that the BrdU positive cells were increased by using 10 and 20 mM calcium MEM, but the positive cells were decreased by using 0 and 2 mM calcium MEM. Increase of alpha-SMA stained cells was recognized when using 0, 10 and 20 mM calcium MEM. The contraction rate of collagen gel was increased by using the 10 and 20 mM calcium MEM. CONCLUSION: The changes of calcium concentration might be an important factor for the development of cataract, posterior subcapsular opacification, and contraction of the lens capsule after cataract surgery.     

The change in immunohistochemical localization of basic fibroblast growth factor (b-FGF) around the lens capsule after extracapsular extraction]

It has been reported that basic fibroblast growth factor (b-FGF) accelerates proliferation of mesenchymal cells and epithelial cells of cornea and crystalline lens in vitro. The immunohistochemical localization of b-FGF on guinea pig lens capsule after extracapsular extraction (ECE) of the lens was studied in order to investigate the role of b-FGF on posterior capsular opacification after ECE and neovascularization. On the 12th postoperative day, immunohistochemical staining of b-FGF was recognized along the posterior capsule on the cortical side, but not along the anterior capsule. On non-operated eyes, scanty immunoreactive materials were seen in cells at the germinative center of the equatorial region. More than two months after ECE, almost no immunoreactive staining could be recognized. In this study, immunoreactive staining of b-FGF was recognized along the posterior capsule on cortical side after operation. It has also been reported that b-FGF stimulates proliferation of the lens epithelial cells in vitro. Therefore, it is suggested that b-FGF plays an important role on posterior capsular opacification after cataract formation.     

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.