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1.
目的观察高血糖对兔眼晶状体摘出术后后囊膜混浊(posterior capsular opacification,PCO)的影响,探讨蛇毒去整合素Kistrin对晶状体上皮细胞(lens epithelial cells,LECs)增殖的抑制作用。方法制作正常血糖及糖尿病兔PCO模型,随机分组:正常血糖组分为A、B两组(每组3只兔3眼),糖尿病组分为C、D两组(每组3只兔3眼),A、C组为对照组(术毕前房注入林格氏液),B、D组为用药组(术毕前房注入80mg·mL-1的Kistrin2mL)。注药后在裂隙灯显微镜下观察4组PCO发生情况及形态并计算增殖指数(PI),注药后14d时取材,检测LECs增生细胞核抗原(proliferating cell nuclear antigen,PCNA)的表达。结果糖尿病兔PCO发生程度重于正常血糖兔;A组PI为0.25±0.05,B组0.03±0.01,C组0.86±0.04,D组0.65±0.08,A组PI明显小于C组(P=0.00),用药后B、D组PI均分别明显小于A、C组(均为P=0.01),但B组PI明显小于D组(P=0.00)。结论与正常血糖兔相比,糖尿病兔的PCO发生率较高,其LECs的增殖能力较强,Kistrin可明显抑制正常血糖及糖尿病兔的LECs增殖。  相似文献   

2.
背景实验性糖尿病动物模型的制作是对糖尿病性眼病进行实验研究的关键环节,目前国内普遍应用的方法是链脲佐菌素和四氧嘧啶(ALX)注射,但前者价格昂贵,后者造模过程中动物的死亡率较高。目的探讨ALX注射制作糖尿病兔晶状体后囊膜混浊(PCO)模型并降低死亡率的方法,并观察高血糖对晶状体PCO形成的早期影响。方法将清洁级健康新西兰雄性大白兔40只随机分为2组;其中20只兔经耳缘静脉一次性注射ALX90mg/kg建立糖尿病模型作为高血糖组,另20只兔以同样的方法注射等量生理盐水作为正常血糖组。药物注射后2周时高血糖组兔血糖升高到12.0mmol/L以上可判断为建模成功,2组分别行兔右眼透明晶状体囊外摘出术并对晶状体PCO进行分级。于术后第6、10、14天取眼球,应用免疫组织化学法观察增生细胞核抗原(PCNA)在后囊膜晶状体上皮细胞(LECs)中的表达情况。结果ALX注射后糖尿病兔的成模率为70%。术后第6、10、14天时,高血糖组兔体质量均明显低于正常血糖组,但血糖明显高于正常血糖组,差异均有统计学意义(P〈0.05)。高血糖组术后14d时观察的3只兔中,2只兔出现后囊膜2级混浊,另1只兔出现1级混浊。正常血糖组术后14d时观察的3只兔后囊膜均出现1级混浊。免疫组织化学染色显示,术后第10天高血糖组可见PCNA在LECs的细胞核中表达,但正常血糖组未见PCNA的表达。术后第14天时高血糖组PCNA增生指数为0.86±0.04,明显高于正常血糖组的0.25±0.03,差异有统计学意义(t=-16.171,P=0.000)。结论90mg/kg的ALX静脉注射能形成稳定的糖尿病兔PCO模型;高血糖是促进PCO发生发展的重要因素之一。  相似文献   

3.
目的 探讨人工晶状体植入术后阿霉素(adriamycin,ADM)抑制兔眼晶状体上皮细胞(rabbit lens epithelial cells,RLEC)增殖的作用及客观评价后囊膜混浊(posteriorcapsule opacification,PCO)的方法。方法 采用兔眼品状体囊外摘出联合人工晶状体植入术的动物模型,术中将携带ADM药物缓释系统植入l5只兔眼晶状体囊袋内。术后观察后囊膜混浊情况;术后第4周,链霉亲和素免疫组化法(strePt—avidin—biotin—enzyme comple,SABC)检测兔眼品状体上皮细胞的增殖细胞核抗原(pro1iferating cell nuclear antigen,PC—NA)的表达。结果 实验组与对照组比较,兔眼晶状体上皮细胞的PCNAP阳性表达下降,2组比较差别具有显著性意义。结论 ADM能有效抑制人工晶状体植入术后RLEC的增殖,免疫组化技术(SABC法)检测RLEC的PCNA的表达,为实验室客观评价PCO提供一种简便有效的方法。  相似文献   

4.
BALB/c小鼠后发性白内障动物模型的建立和观察   总被引:1,自引:0,他引:1  
《眼科新进展》2007,27(2):91-95
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]  相似文献   

5.
目的 观察并探讨不同浓度去整合素(Echistatin,Ecs)对糖尿病兔远期后发性白内障(posteriorcapsularopacification,PCO)发生过程中晶状体上皮细胞(lensepithelialcells,LEC)增殖的抑制作用,以及其在眼内的安全性。方法 建立糖尿病兔模型,随机分为对照组(A组,术毕前房注入蒸馏水0.2mL);实验组:B组、C组、D组(术毕前房分别注入5.0×10-3μgL-1、7.5×10-3μgL-1、10.0×10-3μgL-1Ecs),术后裂隙灯下观察角膜、前房及PCO情况。术后6周取材,常规石蜡切片,HE染色。用免疫组织化学法检测PCO发生过程中LEC增殖细胞核抗原(proliferatingcellnuclearantigen,PCNA)的表达,TUNEL法检测角膜内皮细胞、视网膜神经节细胞层及内核层细胞的凋亡情况。 结果 HE染色结果显示,术后6周各组角膜内皮细胞排列整齐、未见水肿,视网膜内界膜与各层组织结构完整,未见药物引起明显的组织损伤反应。各组间PCO分级差异有统计学意义(P=0.011)。免疫组织化学法检测各组后囊膜PCNA表达结果显示,A组、B组、C组、D组的平均光密度值分别为0.499±0043、0.484±0.055、0.422±0.012、0.340±0.031,B组后囊膜上PCNA表达与A组差异无统计学意义(P>0.05),余组间两两比较差异均有统计学意义(均为P<0.05)。TUNEL法检测各组角膜内皮细胞、视网膜神经节细胞层及内核层细胞凋亡发现,各组间细胞凋亡比较均未见明显差异(均为P>0.05)。结论 10.0×10-3μgL-1Ecs对糖尿病兔远期PCO具有明显抑制作用,且对眼内相邻及重要组织无明显毒性作用,因此可选用此浓度进行后续研究。  相似文献   

6.
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.  相似文献   

7.
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。  相似文献   

8.
目的:探讨γ干扰素对兔后囊混浊发生的防治作用。方法:将30只新西兰白兔60眼随机分为A、B和C三组,三组均行兔晶状体囊外摘除术,A组为对照组,术中晶状体囊袋内灌注BSS;B和C组为治疗组,术中晶状体囊袋内分别灌注106U/L、107U/Lγ干扰素。术后观察晶状体后囊膜变化。3mo后取角膜行光镜、扫描电镜观察角膜内皮变化及晶状体后囊膜铺片HE染色统计晶状体上皮细胞(LEC)的密度,并行免疫组化染色真彩色医学图象分析系统检测增殖细胞核抗原(PCNA)表达积分光密度(IOD)。结果:①3mo后临床观察可见,B组、C组的后囊混浊(PCO)发生率较A组明显减少(P<0.05),C组较B组的发生率少(P<0.05)。统计学分析均有显著性差异。②B组(2972.11±156.47)、C组(2775.23±154.01)后囊膜LEC密度较A组(3297.53±105.60)小,且C组的LEC密度相似文献   

9.
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。  相似文献   

10.
目的 建立一种微创、安全、有效、快速的兔晶状体超声乳化后囊膜混浊模型,为更高要求的实验作准备.方法 新西兰大白兔11只(22只眼),行双眼超声乳化术,分别于术后1 d、3 d、1周、2周和术后1、2、3个月对术眼进行裂隙灯显微镜及组织病理学检查,观察兔眼前、后节反应,后囊膜混浊形成时间、部位、发展过程及组织形态学改变.结果 后囊膜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;术后3个月均出现不同程度的后囊膜混浊.结论 成功建立兔晶状体超声乳化后囊膜混浊模型,可为更高要求研究后囊膜混浊的实验做准备.  相似文献   

11.
目的:研究细胞周期蛋白激酶抑制因子p21和p27蛋白在不同年龄段兔晶状体上皮细胞(lens epithelial cells,LECs)中的表达规律。方法:分别取10,20,30周龄兔晶状体前囊膜组织,采用RT-PCR和Western-blot检测该组织中p21和p27的mRNA水平和蛋白水平。结果:兔晶状体前囊膜组织中p21与p27的mRNA水平和蛋白水平相似,青年最高,老年次之,中年最低。p21和p27在青年、中年及老年兔LECs中的表达水平有差异(P<0.05)。结论:p21和p27在兔LECs周期调控过程中可能起着平衡作用,对防止PCO的研究具有一定参考价值。  相似文献   

12.
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.  相似文献   

13.
目的 观察去整合素echistatin对糖尿病兔后发性白内障形成中信号通路PI3-K/Akt和ERK1/2的影响,从分子水平上探讨echistatin的作用。方法 建立糖尿病兔模型(n=24),随机分组并行透明晶状体囊外摘出术,术毕前房分别注入0.2mL灭菌蒸馏水(对照组,n=12)或0.2mL10.0mg·L-1echistatin溶液(echistatin干预组,n=12)。术后10d及6周(每个时间点6眼),裂隙灯显微镜观察两组术眼PCO分级情况,同时摘取术眼应用RT-PCR法检测后囊膜上信号因子Akt和ERK1/2mRNA表达情况。结果 术后10d对照组和echistatin干预组PCO分级比较差异无统计学意义(P=0.093),但echistatin干预组PCO1级眼数少于对照组;术后6周echistatin干预组PCO级别明显低于对照组(P=0.006)。对照组和echistatin干预组AktmRNA相对表达量术后10d分别为0.9817±0.3804、0.4817±0.1665,术后6周分别为0.6517±0.2047、0.4017±0.1513,echistatin干预组均明显低于对照组(P=0.015,0.037)。对照组和echistatin干预组ERK1/2mRNA相对表达量术后10d分别为0.9483±0.2275、0.6100±0.2806,术后6周分别为0.9217±0.2994、0.4650±0.1800,echistatin干预组均明显低于对照组(P=0.045,0.009)。结论 去整合素echistatin对糖尿病兔后发性白内障的发生和发展有一定的抑制作用,其发生的机制可能与抑制信号因子Akt和ERK1/2表达,进而阻断信号通路PI3-K/Akt和ERK1/2的转导有关。  相似文献   

14.
PURPOSE: To evaluate bag-in-the-lens implantation by studying the feasibility of implanting a new type of intraocular lens (IOL) and the occurrence of posterior capsule opacification (PCO) in human postmortem eyes and in eyes of living rabbits. SETTING: Department of Ophthalmology, University of Antwerp, Belgium, and Netherlands Research Institute of Amsterdam, Amsterdam, The Netherlands. METHODS: The IOL was implanted in 10 postmortem human donor eyes (in vitro study) and in 17 eyes of 10 rabbits (in vivo study). The postmortem capsular bags were cultured for 4 to 6 weeks, and the rabbits were killed 1 to 5 months after implantation. All capsular bags with the bag-in-the-lens were examined by light microscopy and scanning electron microscopy. RESULTS: The IOL design was highly effective in restricting lens epithelial cell (LEC) proliferation in the remaining lens bag in human donor eyes and in rabbit eyes. In eyes in which the capsules were not positioned well within the groove of the IOL, LEC proliferation and PCO occurred. CONCLUSION: Bag-in-the-lens implantation was highly effective in preventing PCO in vitro and in vivo provided the anterior and posterior capsules were secured properly in the peripheral groove of the IOL.  相似文献   

15.
PURPOSE: We have previously shown that patients who have a capsulorrhexis larger than the diameter of a polymethylmethacrylate (PMMA) intraocular lens (IOL) rapidly develop increased posterior capsule opacification (PCO), in effect, producing an example of enhanced PCO. This study focuses on the influence of AcrySof IOLs on this process. METHODS: Phacoemulsification was performed on two groups of patients. The first consisted of 38 patients with a large capsulorrhexis of 6-7 mm who received a 5.5-mm PMMA IOL. The second group of 32 patients had identical surgery and a 5.5-mm MA30 AcrySof IOL was implanted. On days 1,14, 28, 90, 180, and 360, high resolution digitized retroillumination images were taken of the posterior capsule. The PCO area was measured by image analysis at 90, 180, and 360 days. Wrinkling of the posterior capsule was determined at 90 days, and the progression or regression of lens epithelial cell (LEC) proliferation was established by examination of serial images at 28 and 180 days. RESULTS: At 90 days, 79% of the patients with PMMA IOLs had moderate to severe wrinkling of the posterior capsule, whereas the patients with AcrySof IOLs had none (P <.001). The percentage of PCO area was 69% for the PMMA IOLs and 24% for the AcrySof IOL group at 360 days (P <.0001). In the PMMA group, LEC progression occurred in 77%, LEC growth was stable in 15%, and LEC regression occurred in only 8%, compared to 69% of patients with AcrySof IOLs (P <.0001). CONCLUSIONS: In patients with a rhexis larger than the IOL, AcrySof IOLs potentially can prevent capsular wrinkling and cause less PCO than a PMMA IOL with a similar rhexis size. The LEC regression occurs with AcrySof between 28 and 180 days. The reasons for this are discussed.  相似文献   

16.
In vitro model for the study of human posterior capsule opacification   总被引:2,自引:0,他引:2  
PURPOSE: To develop and evaluate a model for the organ culture of human lens capsules that reduces problems inherent in preexisting models for the study of in vitro posterior capsule opacification (PCO). METHODS: Human lenses (N = 110) were isolated from donor eyes and supported externally within a lens holder system by medical-grade cyanoacrylate glue, allowing visualization of the entire capsular bag. After capsulorhexis and lens extraction were performed, the capsule specimens were maintained at physiological conditions for up to 4 weeks. The area of lens epithelial cell (LEC) coverage over the posterior capsule surface was determined objectively on a daily basis using a graticule. Lens epithelial cell behavior was correlated with clinical data and other in vitro PCO models. RESULTS: Cyanoacrylate glue did not appear to be toxic to LECs at the concentration used. The amount of viable epithelium after nuclear extraction was dependent on the age and postmortem time of the specimen. Viable LEC cultures were obtained from eyes up to 9 days postmortem. The time from death to culture or from enucleation to culture did not influence LEC viability if it was fewer than 5 days. The LEC proliferation rates and confluence times were age dependent and correlated closely between pairs of eyes. CONCLUSIONS: Results show that the lens holder model is a more physiological method for supporting the capsule and is a robust, reproducible system for the study of LEC migration and proliferation. It allows visualization within the entire capsular bag. Intraocular lenses can be implanted in this system in a way that more closely resembles the in vivo scenario. This model can be used to evaluate therapeutic measures to prevent PCO.  相似文献   

17.
骆驼蓬总碱及其脂质体对兔晶状体上皮细胞的抑制作用   总被引:9,自引:0,他引:9  
吴明星  利华明 《眼科研究》1999,17(4):275-278
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。  相似文献   

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