摘要
目的观察结缔组织生长因子(CTGF)在视网膜色素上皮(RPE)细胞损伤模型中的表达及其对RPE细胞移行的影响。方法利用体外培养的单层近融合期人RPE细胞,采用棉签和角膜移植用环钻做圆形细胞刮伤区,建立体外RPE细胞损伤模型。链霉亲和素-生物素复合物(SABC)免疫组织化学和原位杂交(ISH)方法检测RPE细胞受创后不同时间CTGF的表达及其转录水平mRNA的变化。并计数进入缺损区的RPE细胞数,定量观察CTGF对RPE细胞移行的影响以及地塞米松对CTGF这一作用的影响。结果免疫组织化学和原位杂交结果显示,创伤后6h,创伤边缘RPE细胞表达CTGF呈弱阳性反应;随着创伤后时间延长,阳性信号逐渐增强;伤后24、48h创伤边缘移行的RPE细胞呈CTGF强阳性表达。重组人CTGF因子(rhCTGF)呈剂量依赖性刺激RPE细胞移行,地塞米松对CTGF诱导的RPE细胞移行作用有显著抑制作用。结论CTGF参与了RPE细胞创伤修复过程,这在增生性玻璃体视网膜病变等眼内增生性疾病中可能有重要意义。
Objective To observe the expression of connective tissue growth factor （CTGF） in injured model of retinal pigment epithelial （RPE） cells and the promoting effect of CTGF on migration of RPE cells. Methods Cultured monolayer-confluent human RPE cells were scraped with a trephine and a cotton stick, and set up the injured model of RPE cells with round scraped area. Immunohistochemistry and in situ hybridization（ISH） were used to detect the expression of CTGF protein and mRNA in injured RPE cells at distinct time points after injury. The number of RPE cells migrated to injured area was measured and the effect of CTGF on migration of RPE cells and the effect of dexamethasone （DEX） on the promoting process of CTGF were observed. Results The results of immunobistocbemistry and ISH indicated the weak positive expression of CTGF in RPE cells at the edge of scrape 6 hours after injury, and the positive expression increased gradually as time goes by after the injury. Strong positive expression of CTGF in RPE cells at the edge of scrape was found 24 and 48 hours after injury. Rebuilt human CTGF stimulated migration of RPE cells in a close-depended manner, and DEX significantly inhabited the migration. Conclusion CTGF involves in the procedure of repair of injury of RPE cells, which may play an important role in the pathogenesis of intraocular proliferative diseases such as proliferative vitreoretinaopathy.
出处
《中华眼底病杂志》
CAS
CSCD
北大核心
2005年第5期306-309,共4页
Chinese Journal of Ocular Fundus Diseases
基金
国家自然科学基金资助项目(30200311)
关键词
内皮生长因子
色素上皮
眼
基因表达
原位杂交
免疫组织化学
疾病模型
动物
Endothelial growth factors
Pigment epithelium of eye
Gene expression
In sita hybridization
Immunohistochemistry
Disease models, animal
