重组人角质细胞生长因子-2对兔碱烧伤模型角膜上皮损伤的治疗作用

J4 ›› 2010, Vol. 36 ›› Issue (4): 625-628.

重组人角质细胞生长因子-2对兔碱烧伤模型角膜上皮损伤的治疗作用

王晓杰^1,2,3,周鑫²,田海山²,马吉胜²,焦悦²,张睿²|蔡敏倩²|钱焕文⁴|李校堃^1,2

1. 吉林大学公共卫生学院卫生部放射生物学重点实验室|吉林长春 |130021；2. 吉林农业大学生物反应器与药物开发教育部工程研究中心|吉林长春 |130118； 3. 温州医学院药学院生物技术制药工程重点实验室| 浙江 温州 325035；4. 军事医学科学院放射与辐射医学研究所|北京 100850

收稿日期:2010-01-27 出版日期:2010-07-28 发布日期:2010-07-28
通讯作者: 李校堃(Tel:0431-84533348,E-mail:xiaokunli@163.net）;钱焕文（Tel:010-66931122,E-mail:laser@nic.bmi.ac.cn) E-mail:xiaokunli@163.net;laser@nic.bmi.ac.cn
作者简介:王晓杰(1972-)|女|吉林省长春市人|副研究员|在读医学博士|主要从事新药的开发与研究。
基金资助:
科技部“重大新药创制”科技重大专项基金资助课题(2008ZX09101-057)；科技型中小企业技术创新基金资助课题（09C26212203306);吉林省长春市科技计划项目资助课题(08YJ45);吉林省长春市企业科技特派员行动计划项目资助课题(09KT03)

Therapeutic effects of kertinocyte growth factor-2 on corneal epithelial wound healing in |rabbit model of alkali burn

WANG Xiao-Jie^1,2,3, ZHOU Xin², TIAN Hai-Shan², MA Ji-Sheng², JIAO Yue², ZHANG Rui², CA Min-Qian², Qian Huan-Wen⁴, LI Jiao-Kun^1,2

1.Key Laboratory of Radiobiology,Ministry of Health,School of Public Health,Jilin University,Changchun 130021,China;2.Engineering Research Center of Bioreactor and Pharmaceutical Development,Ministry of Education, Jilin Agriculture University,Changchun |130118,China；3.Key Laboratory of Biotechnology Pharmaceutical Engineening,School of Pharmacy,Wenzhou Medical College,Wenzhou 325025,China；4.Institute of Radiation Medicine,Academy of Military Medical Sciences, Beijing 100850,China

Received:2010-01-27 Online:2010-07-28 Published:2010-07-28

摘要/Abstract

摘要：

目的: 研究外用旋滴重组人角质细胞生长因子-2（KGF-2）对兔碱烧伤模型角膜上皮损伤的治疗作用，为角膜上皮损伤的早期治疗探索新方法。方法: 制备兔角膜碱烧伤模型，60只日本大耳白兔随机分为碱烧伤对照组和KGF-2组（25 mg/L）(n=30)；各组日本大耳白兔于造模后以50 μL药物滴眼，每日3次。7和14 d裂隙灯观察角膜烧伤面积以及角膜新生血管的形成，MTT法检测角膜上皮细胞活性，免疫组化方法进行病理组织学检查。结果：碱烧伤后7和14 d，KGF-2组角膜烧伤面积小于碱烧伤对照组，差异有显著性（P<0.01）。碱烧伤对照组7 d角膜新生血管发生率为17.50%，KGF-2组为7.32%；碱烧伤对照组14 d角膜新生血管发生率为41.67%，KGF-2组为32.5０%。MTT检测结果显示：碱烧伤后7和14 d，KGF-2组492 nm吸光度与碱烧伤对照组比较差异有显著性（P<0.05或P<0.01）。碱烧伤后7和14 d，病理切片HE染色显示：对照组角膜基质水肿，板层纤维排列松散、紊乱，炎症反应反复出现，角膜上皮层变薄，部分脱落，角膜内皮层内侧有大量成纤维细胞增生；KGF-2组碱烧伤后7 d，角膜上皮细胞生长良好，基质轻度增厚，无明显的炎症反应；碱烧伤后14 d，未出现炎症复发，角膜上皮细胞生长良好，排列整齐，角膜基质仅有轻度增厚。结论：外用旋滴25 mg/LKGF-2可加速角膜上皮损伤的恢复，减轻角膜烧伤的各种炎症反应症状；减轻角膜基质水肿和纤维化，同时KGF-2 亦表现出显著的抑制角膜新生血管形成的作用；KGF-2有可能成为另一个对角膜上皮损伤具有重要调控作用的生长因子。

关键词: 角质细胞生长因子-2；角膜；上皮；损伤修复

Abstract:

To study the therapeutic effect of topically applied keratinocyte growth factor-2 (KGF-2) in vivo on corneal epithelial wound in rabbit model of alkali burn and explore new methods of treatment in the early of corneal epithelial wound. Methods A 6 mm diameter epithelial defect was made in the center of rabbit corneas by the alkali-soaked filter paper.A total of 60 Japanese white rabbits were randomly divided into two groups(n=30): KGF-2 group (25 mg/L) and control group. A 50 μL aliquot of 25 mg/L recombinant human KGF-2 was then applied topically three times a day. The same volume of phosphate-buffered saline (PBS) vehicle was applied to the control.Each corneal epithelial defect was subsequently photographed with a slit lamp on day 7 and 14 after burn and was measured by a computer-assisted digitizer.Corneal neovascularization in two groups was observed every day.The viability of corneal epithelial cells was assessed by MTT assay;meanwhile,the corneal epithelium was examined by morphometric and histological analysis.Results There were statistically significant differences in the epithelial defet area on day 7 and 14 between KGF-2 and control groups(p<0.01).The differences in corneal neovascularization (CNV) between KGF-2 and control groups were also evident on day 7 and 14, the average percentages of CNV in KGF-2- group were 7.32% and 32.50%,whereas the rates in control group were 17.50% and 41.67%,respectively.On day 7 and 14 after wounding,there were statistically significant differences in the mean 570 nm absorbances between KGF-2 and control groups(P<0.05 or P<0.01).Histological examination showed that the corneal lesions consisted of stromal edema,keratocyte loss,inflammatory cell infiltration,and CNV.Corneal epithelium cell desquamation,stromal edema and numerous neutrophil infiltrations were present in the corneal stroma in control group.In contrast,the KGF-2 group exhibited no corneal stroma inflammation,only mild stromal edema remained in KGF-2 corneas on day 14 and the significant changes of the re-epithelialization in KGF-2 group were observed.Conclusion Topically applied 25 mg/L KGF-2 can accelerate corneal epithelial wound healing,in contrast to the control,and reduce inflammation,stromal edema,and fibrosis.In addition,it also exhibites significant inhibition on CNV.KGF-2 maybe become another important growth factor in the regulation of corneal epithelial wound healing.

Key words: keratinocyte growth factor-2;corneal;epithelium;wound healing

中图分类号:

R772.2

王晓杰, 周鑫, 田海山, 马吉胜, 焦悦, 张睿, 蔡敏倩, 钱焕文, 李校堃. 重组人角质细胞生长因子-2对兔碱烧伤模型角膜上皮损伤的治疗作用[J]. J4, 2010, 36(4): 625-628.

WANG Xiao-Jie, ZHOU Xin, TIAN Hai-Shan, MA Ji-Sheng, JIAO Yue, ZHANG Rui, CA Min-Qian, Qian Huan-Wen, LI Jiao-Kun. Therapeutic effects of kertinocyte growth factor-2 on corneal epithelial wound healing in |rabbit model of alkali burn[J]. J4, 2010, 36(4): 625-628.