摘要
目的研究比较改良前后重组人角膜上皮素对人角膜上皮细胞黏附的影响。方法培养人角膜上皮细胞,将野生型角膜上皮素蛋白和诱变型重组角膜上皮素蛋白浓度调整为10,20,30,100μg/mL,分别预孵化96孔酶标板,每种浓度6个孔。包被18 h,孵育2 h。加入人角膜上皮细胞的细胞悬液调整浓度为每孔3×105,每组都设2%BSA预孵化的细胞板对照组。全自动定量绘图酶标仪检测吸光度。结果诱变型重组人角膜上皮素比野生型角膜上皮素拥有更高的促角膜上皮细胞黏附作用,以剂量依赖式方式促进人角膜上皮细胞的黏附。在诱变型重组人角膜上皮素浓度达到10μg/mL时,开始促进人角膜上皮细胞黏附,随着浓度的增高,作用增强;浓度达到30μg/mL时,促进作用达到高峰。结论诱变型人角膜上皮素比野生型角膜上皮素拥有更高的促角膜上皮细胞黏附作用,为进一步临床应用提供了理论基础。
Objective To evaluate the effect of reconstructed human keratoepithelin before and after refining on human corneal epithelial cells adhesion. Methods Human corneal epithelial cells were cultured. Concentrations of wild type keratoepithelin and mutagenized type reconstructed keratoepithelin were adjusted correspondingly to 10,20,30,100μg/mL and used to preincubate 96-hole enzyme label plate, each concentration repeated in six holes for 18 h, culturing for 2 h. Each hole was added human corneal epithelial cells suspension to adjust the concentration to 3×10^5 ceils per hole. Cell plate with 2% BSA preincubation was used as control for each group. Each hole was detected on absorbance using fully automatic quantitative drawing microplate reader. Results Mutagenized type reconstructed keratoepithelin, which worked depending on its dose （takes effect with the concentration of 10 μg/mL, works more with the enhancement of concentration and acts to the peak under the concentration of 30 μg/mL）, was more effective on promoting corneal epithelial cells adhesion compared with wild type keratoepithelin. Conclusion Mutagenized type reconstructed human keratoepithelin is more effective on promoting corneal epithelial ceils adhesion than wild type keratoepithelin, which provides theory basis for clinic application forward.
出处
《哈尔滨医科大学学报》
CAS
北大核心
2009年第4期333-335,共3页
Journal of Harbin Medical University
基金
黑龙江省科技攻关项目(GC08C416)
关键词
角膜上皮素
角膜上皮细胞
黏附
keratoepithelin
corneal epithelial cell
adhesion
