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-  2018 

睫状上皮营养因子对氧化损伤RGC-5的保护作用 Ciliary Neurotrophic Factor Mediated RGC-5 Survival from Oxidative Damage

Keywords: 睫状上皮营养因子,视网膜神经节细胞,神经保护作用,过氧化氢,慢病毒载体

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Abstract:

目的:探究睫状上皮营养因子(CNTF)对过氧化氢所致视网膜神经节细胞(RGC)-5损伤模型的作用。方法:对处于生长对数期的RGC-5予以不同浓度过氧化氢处理不同时间后,根据细胞存活率选择适当的浓度及时间进行造模。造模成功后,予以CNTF进行干预。成功构建携带CNTF基因的慢病毒载体后,将重组慢病毒载体转染的骨髓间充质干细胞(BMSC)与RGC-5共培养。并且由于BMSC可分泌多种神经营养因子,如脑源性神经营养因子(BDNF)、神经胶质细胞源性神经营养因子(GDNF)。RT-PCR检测共培养3d和7d后CNTF和BDNF、GDNF的含量。最后在两种细胞共培养7d后予以过氧化氢损伤,检测细胞存活率。结果:随着过氧化氢浓度及作用时间的增加,细胞存活率随之降低。当过氧化氢浓度为125μmol/L时,达到凋亡高峰,选择该浓度造模。RGC-5与重组慢病毒载体转染的BMSC共培养后,RT-PCR检测发现CNTF和BDNF在培养7d后表达量增多,有统计学意义,而GDNF培养7d后表达量与对照组无统计学差异。当共培养7d后,过氧化氢损伤后检测发现RGC-5存活率增加。结论:CNTF对过氧化氢损伤的RGC-5有保护作用,并且推测在该过程中BDNF同样也发挥了保护作用

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