摘要
目的探讨缺氧条件下促红细胞生成素(EPO)mRNA及蛋白在体外培养的Müller细胞的表达情况。方法胰酶消化新生大鼠视网膜组织制成单细胞悬液,机械震荡、吹打法分离纯化视网膜Müller细胞,反转录-聚合酶链反应(RT-PCR)、免疫细胞化学方法对缺氧条件下视网膜Müller细胞EPO基因和蛋白的表达进行测定。结果成功获得视网膜Müller细胞,传代后95%以上的细胞呈神经胶质纤维酸性蛋白(GFAP)染色阳性。EPO蛋白的阳性染色位于视网膜Müller细胞的细胞浆及突起上。在正常的视网膜Müller细胞中仅见微弱的EPO mRNA和蛋白表达,而缺氧后表达明显上调,且呈时间依赖性。结论Müller细胞在缺氧条件下EPO mRNA和蛋白表达增强,EPO表达水平的升高可能是缺氧性视网膜病变的神经保护因素之一。
Objective To investigate the effect of hypoxia on expressions of erythropoietin（EPO） mRNA and protein in retinal Mueller cells cultured in vitro. Methods Retina tissues from the new-born Wistar rats were dissected into cell suspension after digested by pancreatin. Mueller cells were separated and purified by mechanical concussion and blowing and striking method. The expression of EPO mRNA and protein under the condition of hypoxia was detected by semi-quantitative reverse transcriptase （RT）- polymerase chain reaction （PCR）and immunocytochemical method. Results Retinal Mueller cells were cultured successfully, 95% of which were positively stained by glial fibrillary acidic protein （GFAP）. Positively stained EPO protein was located in the cytoplasm and protuberance. The expression of EPO mRNA and protein was faint in the normal retinal Mueller cells, but increased obviously and timedependently after hypoxia. Conclusion Expression of EPO mRNA and protein increases in Mueller cells after hypoxia,which may be one of the protective factors for the nerves in anoxic retinopathy.
出处
《中华眼底病杂志》
CAS
CSCD
北大核心
2006年第3期196-199,共4页
Chinese Journal of Ocular Fundus Diseases
基金
山东省科技攻关基金资助项目(2004GGB14112)
