缺氧对视网膜Müller细胞中谷氨酸转运体及未折叠蛋白相关因子XBP1、CHOP表达的影响  

Effect of hypoxia on expression of glutamate/aspartate transporter and unfolded protein reaction related factors XBP1,CHOP in retinal Müller cells

作　　者：孙丽娟[1] 胡丹[1] 潘峰[1] 马吉献[1] 苏静波[1] 

机构地区：[1]第四军医大学西京医院眼科,全军眼科研究所,陕西省西安市710032

出　　处：《眼科新进展》2012年第4期305-309,共5页Recent Advances in Ophthalmology

基　　金：国家自然科学基金资助(编号:30772368)~~

摘　　要：目的研究体外缺氧刺激后视网膜Müller细胞是否存在未折叠蛋白反应(unfol-dedprotein reaction,UPR),及其与L-谷氨酸/L-天门冬氨酸转运体(glutamate-aspartatetrans-porters,GLAST)的关系。方法出生3～7d大鼠视网膜组织采用胰蛋白酶消化法培养视网膜Müller细胞,氯化钴(CoCl2)200μmol·L-1对视网膜Müller细胞进行体外缺氧干预,干预时间为0h、3h、6h、12h、24h、48h、72h;UPR诱导剂二硫苏糖醇(DTT)2mmol·L-1刺激视网膜Müller细胞为阳性对照,干预时间为0h、3h、6h、12h、24h、48h、72h。采用实时荧光定量PCR检测GLAST与UPR相关因子X盒结合蛋白1、C/EBP同源蛋白的表达并分析其相关性。结果视网膜Müller细胞鉴定:细胞免疫荧光染色示90%以上细胞GS、GLAST表达阳性;流式细胞仪检测结果:Müller细胞的GLAST表达阳性率为(84.66±3.51)%。缺氧刺激后,GLAST与X盒结合蛋白1、C/EBP同源蛋白在Müller细胞上均有表达,缺氧早期呈上升趋势,干预后24h表达最强,后呈下降趋势。DTT干预组表达趋势与缺氧干预组一致。结论体外缺氧刺激后视网膜Müller细胞存在UPR,其相关因子的变化与GLAST的变化趋势一致,提示UPR可能是调控GLAST变化的原因之一。Objective To investigate the unfolded protein reaction（UPR） related factors X-box binding protein 1（XBP1） and C/EBP homologous protein（CHOP） in retinal Müller cells by hypoxia and the relationship between UPR and glutamate/aspartate transporter（GLAST）.Methods Retinal Müller cells of rat at postnatal 3-7 day were cultured by trypsin-dispersed cell culture.Cultured retinal Müller cells were maintained with Cocl2/DTT for 0 hour,3 hours,6 hours,12 hours,24 hours,48 hours,72 hours in vitro,respectively.The expression of XBP1 mRNA,CHOP mRNA and GLAST mRNA were semi-quantified by Real Time PCR.Results Under hypoxia condition,XBP1,CHOP and GLAST co-expressed in 90% retinal Müller cells,the positive rate of GLAST expression was（84.66±3.51）%.The expression levels of XBP1,CHOP and GLAST were all ascended at the early stage,reached the summit at 24 hours after hypoxia and then dropped,which was equal to the results of DTT.Conclusion UPR exists in the Müller cells after hypoxia,and its related factors XBP1,CHOP expression are equal to GLAST,which suggests UPR may be one cause of the regulation of GLAST changes.

关 键 词：缺氧 未折叠蛋白反应 MÜLLER细胞 GLAST 视网膜 

分 类 号：R339.141[医药卫生—人体生理学]