【摘要】 目的探讨神经营养素-4(NT4)-神经短肽(NAP)对兔虹膜色素上皮细胞(IPECs)的保护作用。方法分离培养兔IPECs,采用非对称互补引物/模板方法构建神经短肽NAP和神经营养因子NT4的融合基因,构建重组腺相关病毒(rAAV)-绿色荧光蛋白(GFP)和rAAV-NAP,并以此转染体外培养的兔IPECs,以荧光表达来检测病毒的转染情况。对体外培养的兔IPECs进行紫外线光损伤,并用MTT法和流式细胞仪来检测NAP对兔IPECs的保护作用。结果转染了rAAV-GFP的兔IPECs表达了明显的GFP荧光。与对照细胞相比,转染了rAAV-NAP的兔IPECs经紫外线光损伤后,细胞的增殖正常,细胞凋亡率降低。结论本研究所构建的rAAV-NAP可有效地转染体外培养的兔IPECs,NAP对兔IPECs有光保护作用。(中华眼科杂志,2006,421008-1012)更多还原

【Abstract】 Objective To investigate the effect of infection with adeno-associated virus (AAV) vector containing NT4-NAP fusion gene on photoprotecion of rabbit iris pigment epithelium cells(IPECs). Methods rAAV-GFP and rAAV-NAP(containg NT4-NAP fusion gene )were constructed; rabbit IPECs were cultured and infected with rAAV-GFP and rAAV-NAP; transfection of viruses was detected by GFP fluorescence expression; NAP protect rabbit iris pigment epithelium from light stimulation was evaluated by MTT and flow cytometry . Results Rabbit IPECs expressed the GFP fluorescence; comparing with control cells, IPECs transfected with rAAV-NAP remained normal proliferation and showed lower apoptosis percentage after ultra-violet stimulation. Conclusion rAAV-NAP constructed in the study can infect rabbit IPECs, and NAP may protect rabbit IPECs from ultra-violet damage.更多还原