摘要
目的探讨豚鼠巩膜成纤维细胞中信号转导及转录活化因子3（STAT3）在胰岛素样生长因子1（IGF-1）诱导中的激活状态及其对细胞的生存及转化作用，明确在模拟近视巩膜细胞中是否存在组成性激活的STAT3信号通路。方法选用也是近视因子的IGF-1刺激豚鼠巩膜成纤维细胞后，免疫化学染色法检测细胞中STAT3及磷酸化STAT3（P-STAT3）蛋白的表达情况，RT-PCR法检测STAT3mRNA的表达；MTT法检测细胞增殖情况。结果细胞免疫化学法显示IGF-1诱导豚鼠巩膜成纤维细胞中存在激活的STAT3信号转导通路，通路蛋白STAT3、P-STAT3在细胞中均有表达（t=-6．925，-10．179，P〈0．01）；半定量RT-PCR结果表明，STAT3mRNA在细胞中也存在过表达（t=9．363，P〈0．01），MTT法显示IGF-1具有促豚鼠巩膜成纤维细胞增殖作用（P〈0．05）。结论IGF-1能诱导体外培养的豚鼠巩膜成纤维细胞中STAT3信号通路的激活，提示STAT3信号通路可能在近视及巩膜重塑发生、发展中起重要作用。
Objective To investigate the expression of signal transducers and activators of transcription 3 （STAT3） in sclera fibroblast of guinea pigs for understanding whether STAT3 signaling transduction pathway induced by the insulin-like growth factor-1 （IGF-1） was constitutively activated. Methods Immunocytochemical staining was used to determine the protein levels of STAT3 and P-STAT3 （ activated STAT3 ） in scleral fibroblasts. RT-PCR was used to detect the mRNA of STAT3. The effects of IGF-1 on scleral fibroblast proliferation were measured by MTr assay. Results Immunocytochemical staining and RT-PCR revealed that STAT3 protein and mRNA were over-expressed in the cells which contained constitutively activated STAT3 signaling transduction pathway （t = -6. 925, - 10. 179 and 9. 363 ; P 〈 0. 001 ）. The results of MTT showed that IGF-1 induced fibroblasts proliferation significantly at concentrations of 0. 5, 1.0, 10. 0 μg/L （ P 〈 0. 05 ）. Conclusion STAT3 signaling transduction pathway is constitutively activated in the treated scleral cells by IGF-1, suggesting that STAT3 signaling transduction pathway may play a critical role in the occurrence of myopia and sclera remodeling.
出处
《中华眼科杂志》
CAS
CSCD
北大核心
2007年第12期1125-1129,共5页
Chinese Journal of Ophthalmology
