角质细胞生长因子促进角膜上皮损伤修复的研究

目的寻找促进角膜上皮损伤修复，治疗持续性角膜上皮缺损的有效方法。方法用３Ｈ－胸腺嘧啶核苷（３Ｈ－ＴｄＲ）掺入及液体闪烁技术，观察不同浓度的外源性角质细胞生长因子（ｋｅｒａｔｉｎｏｃｙｔｅｇｒｏｗｔｈｆａｃｔｏｒ，ＫＧＦ）对体外培养的人角膜上皮细胞生长的影响，由此推算出有效滴眼液浓度并应用于兔眼。用计算机图形分析系统计算角膜上皮愈合速率；用光镜和电镜评估愈合的质量。结果０．１～１００ｎｇ／ｍｌＫＧＦ有明显促进体外培养的人角膜上皮细胞生长的作用（增长率为２７．６６％～７６．７３％），且呈剂量依赖性（ｒ＝０．９２３３，Ｐ＜０．００１）。１μｇ／ｍｌＫＧＦ滴兔眼，加速了角膜上皮损伤修复（愈合速率，ＫＧＦ组为１．７７±０．２３ｍｍ２／ｈ，与对照组１．４９±０．２４ｍｍ２／ｈ比较，Ｐ＜０．０５）。结论外源性ＫＧＦ对体外培养的人角膜上皮细胞有明显的促生长作用，其滴眼液有加速兔眼角膜上皮创伤修复的作用。     

表皮生长因子对晶状体和角膜上皮细胞增生的刺激作用

目的探讨不同浓度的表皮生长因子（ＥＧＦ）对晶状体和角膜上皮细胞增生的刺激作用。方法利用培养的兔晶状体和角膜上皮细胞，通过ＭＴＴ方法，检测细胞增殖密度。培养晶状体上皮细胞的ＥＧＦ浓度（１～２５０ｎｇ／ｍｌ）分为８组；而培养角膜上皮细胞的ＥＧＦ浓度（４～１０００ｎｇ／ｍｌ）分为１０组。结果ＥＧＦ浓度在３２ｎｇ／ｍｌ时，促晶状体上皮细胞的增生作用最强，与无血清组比较差异非常显著（Ｐ＜０．０１）。促角膜上皮细胞增生的最佳浓度为１６ｎｇ／ｍｌ，与无血清组比较差异显著（Ｐ＜０．０５）。结论上述结果为今后在细胞和分子水平上研究白内障和角膜伤口愈合的发生规律及其机制提供实验资料。     

角质细胞生长因子促人角膜上皮细胞生长的研究

目的寻找促进角膜上皮损伤修复的有效方法。方法用３Ｈ胸腺嘧啶核苷（３ＨＴｄＲ）掺入及液体闪烁技术，观察角质细胞生长因子（ＫＧＦ）对体外培养的人角膜上皮细胞ＤＮＡ合成的影响，并计算细胞倍增时间。结果１～１００ｎｇ／ｍｌＫＧＦ有明显促进人角膜上皮细胞ＤＮＡ合成的作用，且呈剂量依赖性（ｒ＝０．９２３３，Ｐ＜０．００１）。１０ｎｇ／ｍｌＫＧＦ明显缩短了细胞倍增时间（３１．５９±４．８８ｈ，与对照组４０．９８±５．２０ｈ比较，Ｐ＜０．０５）。结论外源性ＫＧＦ对体外培养的人角膜上皮细胞有明显的促细胞增生作用。表明ＫＧＦ具有应用于临床，促角膜上皮损伤修复的可能性。     

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。     

碱性成纤维细胞生长因子治疗兔角膜上皮及基质损伤的实验研究

目的 通过动物实验探讨碱性纤维细胞生长因子对角膜上皮细胞、基质细胞损伤的治疗作用。为受损角膜的愈合提供一条新的、有效的途径。方法 将２４只兔全麻下切除角膜前板层,直径８ｍｍ,约１／３角膜厚度。将其随机分为３组,分别滴生理盐水,０．１μｇ／ｍｌ及１．０μｇ／ｍｌ碱性成纤维细胞因子溶液。术后定期用计算机图像处理系统测量角膜损伤后愈合面积,并用放射自显影方法观察角膜基质细胞生长及分布情况。结果 术后１～     

核因子кB在人角膜基质细胞中的表达

目的 探讨核因子кＢ（ＮＦ－кＢ）在人角膜基质成纤维细胞中的基础表达及激活情况。方法 取体外培养的第２或３代人角膜基质细胞,同步化后分为２组：无血清的ＤＭＥＭ液培养的不加药组;含质量浓度为１０μｇ／ｍｌ脂多糖的ＤＭＥＭ液培养６小时的加药组。均采用免疫荧光抗体染色、流式细胞仪计数法和核蛋白提取物凝胶迁移率法检测。结果 流式细胞计数法示,角膜基质细胞内ＮＦ－кＢ的基础表达率为９．４％,脂多糖作用后升高     

多西环素对碱烧伤大鼠角膜组织中IL-1和TNF-α表达的影响

目的　通过建立大鼠角膜碱烧伤模型,研究多西环素对碱烧伤角膜组织中白细胞介素-１（ｉｎｔｅｒｌｅｕｋｉｎ-１,ＩＬ-１）及肿瘤坏死因子-α（ｔｕｍｏｒｎｅｃｒｏｓｉｓｆａｃｔｏｒ-α,ＴＮＦ-α）表达的影响,探索多西环素作用于角膜碱烧伤的抗炎机制。方法　使用自制模具建立６４只ＳＤ大鼠一侧眼角膜碱烧伤动物模型,分成以３ｇ?Ｌ－１多西环素眼液滴眼的实验组和以溶媒滴眼的阴性对照组,每日滴眼４次。于碱烧伤后３ｄ、７ｄ、１４ｄ、２１ｄ分别取下３２只大鼠烧伤侧角膜组织行酶联免疫吸附实验（ＥＬＩＳＡ）,检测角膜组织中ＩＬ-１、ＴＮＦ-α的表达情况。结果　ＥＬＩＳＡ检测显示碱烧伤后第３天阴性对照组ＩＬ-１蛋白相对表达量为（１２７．５８±３５．９２）μｇ?ｇ－１,第７天为（８９．５４±４２．６３）μｇ?ｇ－１,第３天多西环素组ＩＬ-１蛋白相对表达量为（５３．７７±２１．１８）μｇ?ｇ－１,第７天为（３６．６０±１０．７８）μｇ?ｇ－１,第３天、第７天时多西环素组ＩＬ-１与阴性对照组相比表达显著下降（均为Ｐ＜０．０５）;第１４天、第２１天与阴性对照组比较ＩＬ-１蛋白表达差异均无统计学意义（均为Ｐ＞０．０５）。碱烧伤后第３天阴性对照组ＴＮＦ-α蛋白相对表达量为（６９．４５±２１．４３）μｇ?ｇ－１,第７天为（５５．３７±１８．３４）μｇ?ｇ－１,第３天多西环素组ＴＮＦ-α蛋白相对表达量为（２４．０６±９．３９）μｇ?ｇ－１,第７天为（１４．８１±５．７２）μｇ?ｇ－１,碱烧伤后第３天、第７天时多西环素组ＴＮＦ-α与阴性对照组相比表达显著下降（Ｐ＜０．０５）;第１４天、第２１天与阴性对照组比较ＴＮＦ-α蛋白表达差异均无统计学意义（均为Ｐ＞００５）。结论　多西环素作用于碱烧伤大鼠角膜可下调ＩＬ-１、ＴＮＦ-α基因的表达,这可能是多西环素下调炎症因子表达,减轻炎症反应的机制之一。     

眼表重建术后局用FK-506及rhEGF临床观察

目的：寻找眼表重建术后主要并发症的有效治疗措施。方法：对３２例３２只严重化学伤患眼施眼表重建术（板层角膜移植、角膜移植羊膜移植术）,按随机方法将研究对象分为治疗组及对照组各１６眼;治疗组于术后一周局部应用０．５ｍｇ／ｍｌ的ＦＫ－５０６及２０ｕｇ／ｍｌ的ｒｈＥＧＦ滴眼液,对照组应用１％ＣｓＡ及Ｓｏｌｃｏｓｅｒｙｌ Ｅｙｅ Ｇｅｌ,观察两组移植组织的排斥反应及上皮细胞愈合质量。结果：在随访期内,治疗组     

重组人表皮生长因子促进角膜上皮损伤修复的研究

目的：客观评估重组人表皮生长因子（ｒｈＥＧＦ）促进角膜上皮损伤修复的临床疗效及安全性。方法：应用前瞻性评估研究方法，对于角膜缘干细胞异常的角膜盲１５３例１５３只眼施行穿透性角膜移植术。移植片直径流一为７．５ｍｍ，术中去除供体角膜上皮；将所有研究病例按随机双盲原则分试验组（９１眼）、赋型剂组（３１眼）及素高捷疗组（３１眼）。术后分别观察移植片角膜上皮的愈合速率。结果：各ｒｈＥＧＦ组术后移植片角膜上皮     

氟喹诺酮类药物抗单纯疱疹病毒1型的实验研究

采用细胞培养法，试验了氟喹诺药物对眼科常见病病毒－－单纯疱疹病毒１型（ＨＳＶ－１）的作用。本类药物对细胞毒性研究表明，氧氟沙星对细胞半数抑制浓度（ＩＤ５０）最高毒性最低。其次为洛美沙星、诺氟沙星、依诺沙星、环丙沙星。五种氟喹经物抗ＨＳＶ－１的半数有效浓度（ＥＤ５０）分别为氧氟沙星１００μｇ／ｍｌ、洛美沙星１００μｇ／ｍｌ、诺氟沙星１４３．７μｇ／ｍｌ、环丙沙星７０．７μｇ／ｍｌ、依诺沙星７０．７μ     

翼状胬肉术后不同处理方法对创面上皮修复的影响

目的：比较包扎术眼及局部滴用重组人表皮生长因子对翼状胬肉术后角膜上皮修复的影响。 方法：单侧原发性翼状胬肉38例38眼,胬肉侵入角膜在2~5mm范围内。均行翼状胬肉单纯切除术,术后将病例随机分为两组,A组19例术后每日结膜囊内涂氧氟沙星眼膏,绷带包眼至角膜上皮完全愈合;B组19例术后当日给予结膜囊内涂氧氟沙星眼膏包眼,自术后第2d起每日给予氧氟沙星眼水4次+重组人表皮生长因子眼水4次滴眼。对两组病例的角膜情况进行观察,统计角膜上皮完全愈合的时间,进行比较分析。 结果：角膜上皮平均愈合时间A组为4.1±1.4d,B组为65±2.9d,两组间比较有显著统计学差异（P<0.01）。 结论：翼状胬肉术后包眼较局部滴用表皮生长因子眼水更能加快角膜上皮愈合。     

rhEGF基因治疗角膜碱烧伤的实验研究

目的探讨重组人表皮生长因子（rhEGF）基因缓释型滴眼液对免角膜碱烧伤的基因治疗。方法新西兰白兔75只，分为A（rhEGF-pcDNA3．1-脂质体基因滴眼液）、B（rhEGF衍生物滴眼液）、c（pcDNA3.1空质粒-脂质体对照组）三组分别点眼治疗免角膜碱烧伤。每天观察模型眼眼表、荧光素染色并照相。分别测定hEGFmRNA、hEGF和EGFR在角膜组织中的表达。结果A组眼表结膜充血、分泌物、角膜水肿和角膜混浊程度均比B组和C组轻。角膜荧光素染色显示A组角膜上皮的溃疡面积较B组和C组范围小，21d转为阴性。A组基因在行滴眼24h后在全层角膜细胞内可见hEGFmRNA和蛋白质表达，第3d达高峰，细胞阳性率高达95％，随后逐渐降低，21d仍有弱表达。结论rhEGF-PcDNA3．1-脂质体基因滴眼液能够降低角膜的炎症反应，促进角膜上皮细胞的增生，加速角膜碱烧伤的损伤修复过程。     

Effects of autologous serum eye drops on corneal wound healing after superficial keratectomy in rabbits

PURPOSE:: To study the effects of topically applied autologous serum on the wound healing process of mechanically produced corneal ulcers. METHODS:: A superficial keratectomy was done in 1 eye from each of 28 rabbits, using a corneal trephine. Four groups of 7 rabbits were treated topically 4 times a day with nondiluted autologous serum; diluted autologous serum (20%); unpreserved pharmaceutical tear substitute; and physiologic saline. The saline-treated group served as a control. The healing of the ulcers was followed on a slit-lamp biomicroscope regarding its size, infiltration, and neovascularization on alternate days up to day 10 with and without fluorescein staining. RESULTS:: Autologous serum significantly accelerated the corneal wound healing process in both concentrations compared with either the unpreserved pharmaceutical tear substitute or physiologic saline solutions, but the process was faster with the nondiluted concentration. CONCLUSION:: Autologous serum eye drops could be used as a corneal wound healing adjuvant in mechanical corneal ulcers. To accelerate the wound healing process, the blood-derived factors were brought externally to the avascular cornea through autologous serum eye drops. The importance of the vascular supply on the wound healing process of vascular tissues is well known, and this study considers the benefit of blood-derived factors on the healing process of cornea.     

角质细胞生长因子2促进兔角膜碱烧伤上皮愈合机制的研究

目的　探讨角质细胞生长因子2 (KGF- 2 )对实验性兔角膜中央碱烧伤后角膜上皮愈合的作用及其机制。方法　24只新西兰白兔的24只角膜碱烧伤眼按随机数字法分成4组,每组6只眼,其中A、B、C组为治疗组,分别以3种不同浓度: 1μg/ml、50μg/ml、100μg/mlKGF 2滴眼液治疗;D组为对照组,用磷酸盐缓冲液(PBS)滴眼液治疗;观察角膜上皮愈合情况,并做形态学检查及P63、角质蛋白单克隆抗体(AE5)、表皮细胞生长因子单克隆抗体(EGFR)免疫组化研究。结果　1 ~100μg/mlKGF 2能够促进兔角膜上皮愈合。角膜碱烧伤24h后100μg/mlKGF- 2组和对照组角膜上皮愈合率分别为40%和74% (P<0 .05);第4天时4组均出现一定反复;第10天时各治疗组近完全愈合。碱烧伤后第7天即可观察到P63阳性细胞不仅存在于角膜缘区的部分基底细胞中,同时也向角膜内迁移。如角膜碱烧伤后第7天角膜缘区P63阳性细胞数: 100μg/mlKGF 2组为( 53 .8±2. 6)个,对照组为(29. 5±2. 2)个,正常角膜为(17. 0±2. 1)个(P=0. 000);同时非角膜缘区P63阳性细胞数分别为: 100μg/mlKGF 2组为(69. 5±2. 8)个,对照组为(19 5±2 8)个,正常角膜为0个(P=0 .000)。结论　KGF- 2可激活角膜上皮干细胞,使其不断增殖分化,从而促进角膜上皮损伤的愈合。     

重组人表皮生长因子治疗外伤性角膜上皮缺损

目的　评价重组人表皮生长因子 (rhEGF)滴眼液治疗外伤性角膜上皮缺损的临床疗效。方法　观察 84例外伤性角膜上皮缺损病例 ,随机分成rhEGF滴眼液治疗组和常规治疗对照组 (0 3 %氧氟沙星滴眼液 )各 42例。治疗组用rhEGF滴眼液 ,对照组用 0 3 %氧氟沙星滴眼液 ,每日滴眼 4次 ,共 10天。隔日观察上皮缺损修复情况。结果　rhEGF治疗外伤性角膜上皮缺损的有效率明显高于常规治疗对照组 ,两者差异有非常显著意义 (P <0 0 1)。结论　rhEGF滴眼液治疗外伤性角膜上皮缺损能有效地促进角膜上皮缺损的修复。     

角膜碱烧伤bFGF滴眼治疗作用的形态定量研究

目的 探讨兔角膜碱烧伤后碱性成纤维细胞生长因子(bFGF)对角膜修复和新生血管形成的影响。方法 制作兔角膜碱烧伤模型,烧伤后应用bFGF滴眼液(A组 bFGF浓度为2400ng/ml,B组为9600ng/ml,C组为对照组),以计算机图像分析系统定量测定角膜上皮愈合面积和上象限角膜新生血管面积。结果 图像分析测得角膜上皮愈合面积实验组比对照组大,在第5、9、14 d A组与对照组比较差异有显著意义(P<0.05),在30d B组与对照组差异有显著意义(P<0.05)。图像分析测得上象限角膜新生血管面积实验组与对照组比较差异无显著意义(P>0.05)。结论 bFGF可以促进角膜上皮修复愈合,未见促进角膜新生血管形成等有害作用。     

Corneal wound healing after penetrating keratoplasty with EGF application. Experimental studies

PURPOSE: To check post penetrating keratoplasty (PK) corneal wound healing characteristics after epidermal growth factor (EGF) application and to compare it with controls. MATERIAL AND METHODS: The PK was performed in the group of 72 young, healthy New Zealand rabbits (36 females and 36 males). Slit-lamp examination, tonometry and corneal topography by Tomey's corneal modeling system (TMS-1) were carried out before and after surgery. The PK was performed in both eyes. Half of animals were used as a bilateral donor for the other half, with a rule: right eye cornea to the right eye and left eye cornea to the left eye. As a result, after completed surgery 36 rabbits had bilateral grafts. The animals were divided into 3 equal groups (12 in each). Two drops of the human recombined EGF dissolved in the saline solution with concentration varied from 500 to 1500 ng in each drop were applied to the right eye according to schedule. The left eye was used as a control and did not receive EGF. Time of observation varied from 24 hours to 6 months. The tensinometry and the histopathologic study--light and electron microscopy were performed to determine corneal scarring. RESULTS: The wound healing pattern after PK was characteristic and constant in each group. The corneal wound healing significantly accelerated in the EGF treated group of rabbits compared with the controls (p < 0.05). In the group of rabbits receiving 1000 ng of hrEGF 3 times/day, after two weeks of application we noted increase of the wound strength up to 600 folds, comparing with controls. Well-organized scar was histologically seen on the 21st post-surgery day. The post-operative corneal astigmatism was less expressed in the eyes treated with EGF comparing to controls. CONCLUSIONS: These preliminary results of our experimental study indicated accelerated effect on the corneal wound healing after PK with topical, low dose hrEGF application. Clinical observation of utilization of similar low doses of the hrEGF after PK--is in progress.     

Topical vitamin C,vitamin E,and acetylcysteine as corneal wound healing agents: A comparative study

Purpose:The purpose of this study was to evaluate wound healing abilities and efficacy of topical Vitamin C, Vitamin E, and acetylcysteine for their possible clinical use.Methods:The study was conducted on 36 eyes of 18 single-breed rabbits, Oryctolagus cuniculus, of both sexes. A 7.5 mm calibrated vacuum corneal trephine was used to induce a defect of 100 micron depth in center of both the corneas. The right eye of rabbits was selected as the control eye and the left eye as the trial eye, which received eyedrops as Group 1–10% Vitamin C eyedrops, Group 2–3% acetylcysteine eye drops, and Group 3–1% Vitamin E eye drops. Control eyes received ringer lactate. Evaluation was done for fluorescein stain positivity, epithelial defect size, and corneal haze on Day 2, Day 7, and Day 14 post induction of the epithelial defect.Results:On day 14, three eyes of control group, all Vitamin C and acetylcysteine treated eyes, and four Vitamin E treated eyes were fluorescein stain negative. The mean defect area on day 14 was 0, 0, 0.13, and 1.86 mm² in Vitamin C, Vitamin E, acetylcysteine, and control eyes, respectively. Vitamin C and Vitamin E control corneal haze better than acetylcysteine in experimentally induced corneal wounds in rabbits.Conclusion:The three trial drugs with different mechanism of action showed similar effect on healing of the experimentally created corneal wounds in rabbits, with comparison showing statistical insignificance.     

重组人表皮生长因子滴眼液对白内障超声乳化术后角膜水肿的影响

目的 观察重组人表皮生长因子(recombinant human epidermal growth factor,rhEGF)滴眼液对老年性白内障超声乳化术后角膜内皮水肿发生程度的影响.方法 采取随机对照试验,试验组56例(70只眼),手术前后均应用rhEGF滴眼液;对照组53例(62只眼),除不使用rhEGF滴眼液外,术前术后用药均与试验组相同,观察术后1、3、7 d角膜水肿程度及术后1、7 d中央角膜厚度.结果 试验组术后1、3、7 d角膜水肿程度均低于对照组,差异有统计学意义(P<0.05),两组术后1、7 d中央角膜厚度差异有统计学意义(P<0.05).结论 老年性白内障患者超声乳化手术前后使用rhEGF滴眼液降低了角膜水肿的发生率,减轻了角膜水肿的严重程度.