pEGFP/Ang-1转染BMSCs对高糖环境中RF/6A细胞的保护作用

pEGFP/Ang-1 transfected BMSCs attenuates glucose-damaged RF/6A cells

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。

Abstract:

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 co-cultured 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.