TGF-β_2对豚鼠实验性近视眼后极部巩膜成纤维细胞的增殖和超微结构的影响

The Effect of Transforming Growth Factor-β_2 on Cell Multiplications and Ultra-structures of Posterior Scleral Fibroblasts in Guinea Pigs with Myopia

目的明确转化生长因子β2(TGF-β2)对豚鼠镜片诱导型近视眼后极部巩膜成纤维细胞的增殖和超微结构的影响。方法选择2周龄豚鼠10只,随机选取一只眼(实验组)用镜片诱导法制备近视动物模型,对侧眼为自身对照组。造模30 d后对两组进行屈光度和眼轴检测。培养豚鼠后极部巩膜成纤维细胞。将每只眼培养的细胞分为空白对照及TGF-β21、10、100 ng/ml浓度组,用四甲基偶氮唑盐比色法(MTT)检测各组光吸收值(OD)和细胞增殖的程度;利用电镜观察TGF-β210 ng/ml时细胞的超微结构并对结果进行统计分析。结果凹透镜诱导后实验组屈光度和眼轴与诱导前比较差异均有统计学意义(P<0.05);实验组和自身对照组TGF-β2 100 ng/ml时OD值与空白对照比较差异无统计学意义(P>0.05),TGF-β2 1、10 ng/ml和空白对照两两比较差异有统计学意义(P<0.05),同浓度TGF-β2组间OD值和增殖率比较差异无统计学意义(P>0.05)。不同浓度TGF-β2时细胞增殖率组内比较差异有统计学意义(P<0.05)。在10 ng/ml TGF-β2作用下巩膜成纤维细胞核外形不规则,粗面内质网扩张,细胞器减少,细胞界膜不清,部分界膜消失。结论 TGF-β2可有效地抑制豚鼠巩膜成纤维细胞的增长,其中10 ng/ml的刺激作用最强,可使细胞的超微结构发生变化。

Objective To investigate the effect of transforming growth factor-β2（TGF-β2） on cell multiplications and ultra-structures of posterior scleral fibroblasts in guinea pigs with myopia.Methods Ten 2-week-old guinea pigs were chosen,and concave lens was used to induce myopic models（experiment group）,while the other eye of the same guinea pig serve as a self control group.After modeling for 30 days,the dioper and axis oculi in both groups were detected,and scleral fibroblasts of each group were cultured.The scleral fibroblasts cells were divided into four groups： group A（blank control）,group B（TGF-β2 1 ng/ml group）,group C（TGF-β2 10 ng/ml） and group D（TGF-β2 100 ng/ml）.Optical density（OD） and cell multiplication were assayed by MTT and ultra-structure in group C was studied by electron microscope.Results Diopter and axis oculi in the two groups showed significant difference after concave lens induced（P〈0.05）.There was no significant difference of OD value between group A and D（P〉0.05）,but there was significant difference between group A,B and C（P〈0.05）.There were no significant differences of OD value and inhibition effect between experiment group and self control group（P〉0.05）,but cell multiplication rate showed significant difference in the same group for different concertrations of TGF-β2（P〈0.05）.Posterior scleral fibroblasts in group C showed irregular shapes,dilatedrough endoplasmic reticulum,increased organelles,and unclear limiting membrane or even partly disappeared.Conclusion TGF-β2 can effect the proliferation of scleral fibroblasts cells in guinea pigs,the concentration of 10 ng/ml is the best,which may induce ultra-structure changes.