摘要
目的探讨酮咯酸氨丁三醇（Ketorolac tromethamine 0.5%, Acular,安贺拉）对培养兔角膜细胞及细胞外基质纤连蛋白（Fibronectin,FN）的影响。方法选用原代及传代兔角膜细胞,实验组加入含不同浓度酮咯酸氨丁三醇的培养液,对照组加入等量空白培养液,采用四甲基偶氮唑蓝（MTT）比色法检测其对细胞增殖的抑制,倒置相差显微镜观察其对细胞的形态学改变,并进一步采用免疫细胞化学和ELISA研究其对角膜细胞分泌FN的影响。结果酮咯酸氨丁三醇浓度为50-175μg/ml时,作用24、48、72h均抑制角膜细胞的增殖（P均〈0.05）,并呈剂量依赖性。免疫细胞化学显示对照组可见胞浆着色及胞间丝状着色,而加药组仅见胞浆少量着色。ELISA结果显示低浓度时FN含量增加,而高浓度时其含量明显减少。结论酮咯酸氨丁三醇对角膜细胞的增殖有抑制效应,呈剂量依赖性,并且高浓度时可明显减少FN的含量。酮咯酸氨丁三醇可能通过抑制角膜细胞的增殖和减少细胞外基质,从而防止屈光术后的屈光回退。
Objective To study ketorolac tromethamine for the effect of rabbit keratocytes and the fibronectin. Methods The primary culture and subculture of were exposed to different concentrations of ketorolac tromethamine in experimental group and to culture media in contrast group. The inhibtive characteristics of ketorolac tromethamine to the cells＇ proliferation were measured with methyl thiazolyl thiazolium （MTT） assay. The morphology of cells were observed with light microscope. The changes of the fibronectin were measured by immunocytochemistry. And an enzyme-linked immunosorbent assay（ELISA） was used to determine the content of the fibronectin in supernate. Results After the treatment of 50-175μg/ml ketorolac tromethamine, rabbit keratocytes showed the marked inhibition whenever in 24h, 48h or 72h （P〈0.05） and showed dosage dependence. Immunocytochemistry showed the positive staining of fibronectin was sronger in cell covers of control group than that of experimental group. ELISA method demonstrated increase in supernate FN content of low concentration while decrease it of high concentration. Conclusion Ketorolac tromethamine has obvious inhibitive effect on rabbit keratocytes and showed dosage-effect relation.The supernate FN content was decreased obviously in high concentration group, Ketorolac tromethamine likely lightens the regression of refractive surgery by means of antiproliferative effect and reducing the extracellular matrix.
出处
《中国实用眼科杂志》
CSCD
北大核心
2007年第5期552-555,共4页
Chinese Journal of Practical Ophthalmology
基金
本课题为重庆市卫生局科技计划项目资助项目（04-2-072）.
