摘要
目的:观察血管生成素-1/重组质粒(pEGFP/Ang-1)转染的大鼠骨髓间充质干细胞(BMSCs)对高浓度葡萄糖损伤猴脉络膜-视网膜内皮细胞(RF/6A)的保护作用。方法:以pEGFP/Ang-1转染BMSCs,倒置荧光显微镜下观察增强型绿色荧光蛋白的表达,再利用Transwell模型,将转染的BMSCs与RF/6A共培养于高浓度葡萄糖培养基中。3d后MTT法检测RF/6A活力,Western blot检测磷酸化蛋白激酶B(phosphorylated protein kinase B,P-PKB)的表达,从而探讨转染pEGFP/Ang-1的BMSCs对高浓度葡萄糖培养中的RF/6A的保护作用。结果:成功转染pEGFP/Ang-1的BMSCs可见增强型绿色荧光蛋白表达,与转染pEGFP/Ang-1的BMSCs共培养的RF/6A细胞活力及P-PKB的表达均高于未转染组(均P<0.01),与对照组无明显差别(均P>0.05)。结论:质粒pEGFP/Ang-1转染的BMSCs对高糖环境中的RF/6A具有保护作用,其机制可能与P-PKB表达上调有关。
AIM：To investigate the protective effects of pEGFP/Ang-1 transfected BMSCs on RF/6A in condition of high glucose concentration.
METHODS, BMSCs were transfected pEGFP/Ang-1, confirming by observation of enhanced green fluorescent protein （EGFP） with fluorescence microscope. Rhesus chorioido-retinal endothelial cells （RF/6A） were cultured with or without pEGFP/Ang- 1 transfected BMSCs in transwell under high glucose concentration. Three days after co-cultured, cell vitality and pPKB expression of RF/6A were detected with methyl thiazolyl tetrazolium （MTT） methods and Western blot respectively, RESULTS. EGFP was expressed in BMSCs successfully transfected with pEGFP/Ang- 1. RF/6A cocultured with pEGFP/Ang- 1 transfected BMSCs showed higher viability and pPKB expression compared with RF/6A cultured in condition of high glucose concentration cultured with or without untransfected BMSCs （ P〈 0.01 respectively）, showing no difference to control group （ P〉 0.05 respectively）. ~ CONCLUSION- pEGFP/Ang- 1 transfected BMSCs showed protective effects, may via PKB, on RF/6A in condition of high glucose concentration. KEYWORDS： angiopoietin-1 ; bone marrow mesenchymal stem cells; Rhesus choroidal-retinal vascular endothelial cell; high glucose concentration
出处
《国际眼科杂志》
CAS
2014年第5期807-810,共4页
International Eye Science
基金
国家自然科学基金青年基金(No.81300931)~~
