胶质细胞谷氨酸转运体-1a反义寡核苷酸的设计及效果评价  

Designation and evaluation of antisense oligodeoxynucleotides targeted to glial glutamate transporter-1a

作　　者：刘丽哲[1] 张敏[1] 刘宜先[2] 崔鑫[1] 胡玉燕[1] 李文斌[1] 

机构地区：[1]河北医科大学病理生理学教研室 [2]河北医科大学生理学教研室,石家庄050017

出　　处：《中国应用生理学杂志》2015年第3期238-243,共6页Chinese Journal of Applied Physiology

基　　金：国家自然科学基金资助项目(81271454)

摘　　要：目的:应用IDT公司的Antisense design software设计胶质细胞谷氨酸转运体-1a(GLT-1a)的反义寡核苷酸(AS-ODNs),并对其进行效果评价。方法:首先根据GLT-1a mRNA羧基末端特异序列,设计GLT-1a AS-ODNs;在此基础上,应用Western blot方法对设计的5条GLT-1a AS-ODNs抑制大鼠海马CA1区GLT-1a蛋白表达的效果进行评价。结果:序列为5’-GGTTCTTCCTCAACACTGCA-3’的GLT-1a AS-ODNs可特异性地抑制大鼠海马CA1区GLT-1a蛋白表达,而对GLT-1b的表达无影响。该AS-ODNs对sham和头孢曲松钠(Cef)诱导的大鼠GLT-1a的表达上调的抑制效果类似,均>60%;对脑缺血预处理(CIP)诱导的大鼠海马CA1区GLT-1a蛋白表达的上调亦具有明显的抑制作用,抑制效果>60%。结论:从设计的5条GLT-1a AS-ODNs中获得了可特异性抑制GLT-la表达的反义寡核苷酸序列。Objective： The present study was undertaken to design antisense oligodeoxynucleotides （AS-ODNs） of glial glutamate trans- porter-la （GLT-la） and to evaluate the effectiveness of the designed AS-ODNs on the expression of GLT-la. Methods： Five sequences of GLT-la AS-ODNs were designed according to the C terminus specific sequences of GLT-la mRNA using antisense design software of IDT Com- pany. Western blot analysis was used to evaluate the inhibition effects of the five GLT-la AS-ODNs on the expression of GLT-la. Results： The sequence of GLT-la AS-ODNs with sequence of 5＇ -GGTFCTI＇CCTCAACACTGCA-3＇ could specifically inhibit the expression of GLT-la in the hippocampal CA1 subfield of rats, while it had no effect on the expression of GLT-lb. This sequence showed similar inhibition on the expres- sion of GLT-la in sham and ceftriaxone （Cef） -treated rats. It could also significantly inhibit the cerebral ischemic preconditioning （CIP） -in- duced up-regulation in the expression of GLT-la. The magnitude of the inhibition in sham, Cef- or CIP- treated rats was similar by more than 60%. Conclusion： From the designed five sequences of GLT-la AS-ODNs, we obtained an effective sequence which can specifically inhibit the expression of GLT-1a.

关 键 词：GLT-1a 反义寡核苷酸 头孢曲松钠 脑缺血预处理 海马 大鼠 

分 类 号：R363[医药卫生—病理学]