TGF-β2对豚鼠实验性近视眼后极部巩膜成纤维细胞MMP-2表达的影响

摘要

目的 明确转化生长因子β2(transforming growth factor-β2,TGF-β2)对豚鼠镜片诱导型近视眼后极部巩膜成纤维细胞基质金属蛋白酶(matrix metallo proteinase,MMPs)表达的影响.方法 选择2周龄已脱离母乳喂养的豚鼠12只,随机选取12只中各一眼用镜片诱导法制备近视动物模型(实验组),对侧眼不予任何处理(自身对照组).30 d后采用组织块培养法培养豚鼠后极部巩膜成纤维细胞,并传2代.将每只眼(两组24只眼)培养出的细胞分别分为空白对照组、1 ng/ml TGF-β2组、10 ng/ml TGF-β2组、100 ng/ml TGF-β2组,利用实时荧光定量聚合酶链反应、western-blot蛋白印迹法对各组细胞MMP-2、MMP-2mRNA进行定量分析.结果 经30 d镜片诱导,实验组诱导出(-6.70±1.93)D的相对近视,眼轴相对延长(1.53±0.31)mm,实验前后比较差异有统计学意义(P＜0.05).实验组中空白对照组与自身对照组中空白对照组比较,MMP-2、MMP-2mRNA表达量增高,差异有统计学意义(P＜0.05).TGF-β2浓度为10 ng/ml时实验组和自身对照组细胞MMP-2、MMP-2mRNA的表达量最高,与本组其他浓度比较差异有统计学意义(P＜0.05).结论 凹透镜诱导可引起豚鼠明显轴性近视,TGF-β2能有效刺激MMP-2的表达.%Objective To identify the effect of TGF-β2 on the expression of MMP-2 in posterior scleral fibroblasts in enriettiis with lens-induced myopia. Methods 12 two-week-old enriettiis not dependent on breast milk were selected. One side of eye in all the animals was randomly chosen to be induced as myopia models by lens (experiment group), and the other side without any treatment was regarded as autologous control group. 30 days later, the scleral fibroblasts in both groups were cultured and passaged 2 generations in vitro. The cultured cell of chosen eyes (a total 24 eyes in the two groups) was divided into blank control group; 1 ng/ml TGF-β2 group, 10 ng/ml TGF-β2 group, and I00 ng/ml TGF-β2 group. MMP-2, MMP-2 mRNA were quantitated by PCR fluorometric analysis and western-blot immunoblotting. Results 30 days after lens inducing,when compared with that of control group, relative myopia in experiment group was induced to ( - 6. 70 ± 1.93) D, and axis oculi was relatively prolonged to (1.53 ± O. 31 )mm, and there were significant differences in experiment group before and after the experiment (P ＜ O. 05 ). Compared with blank control group of experiment group, expressions of MMP-2, MMP-2mRNA were higher in the blank control group of autologous control group, and the differences were significant (P ＜ O. 05). Compared with other concentrations of TGF-β2, the expressions of MMP-2, MMP-2mRNA were highest in all groups when the concentration of TGF-β2 was 10 ng/ml, and the differences were significant (P ＜0. 05). Conclusiou Concave lenses can induce significant axial myopia in young Enriettiis, and TGF-β2 may stimulate the expression of MMP-2 effectively.