脱氧核酶对Period1基因表达及其对小鼠吗啡精神依赖的影响  

Inhibitory Effects of Deoxyribozyme on the Expression of Period1 Gene in vitro and on Morphine-induced Psychic Dependence in Mice

作　　者：周薇[1] 刘延友[1] 王跃 汪宇錡 肖静[1] 朱彬[1] 彭文珍[2] 王正荣[1] 

机构地区：[1]四川大学华西基础医学与法医学院生物医学工程研究室 [2]四川大学华西基础医学与法医学院生物化学与分子生物学教研室

出　　处：《四川大学学报（医学版）》2006年第5期734-737,共4页Journal of Sichuan University(Medical Sciences)

基　　金：国家自然科学基金(批准号39970275;30070278)资助

摘　　要：目的研究脱氧核酶在细胞水平调节Period1基因表达的作用及注射脱氧核酶对小鼠吗啡精神依赖的影响。方法设计合成Period1mRNA的10-23型脱氧核酶(DRz164),将pcDNA3-Per1和DRz164在脂质体介导下转染NIH3T3细胞,通过逆转录-聚合酶链反应(RT-PCR),流式细胞术(FCM)检测转染细胞内Period1基因表达水平。脑室注射DRz164,建立小鼠条件位置偏爱模型,观察对吗啡精神依赖的影响。结果转染DRz164后,RT-PCR半定量检测Period1mRNA与对照组相比减少42.4%。细胞内PER1蛋白表达减少57.2%。给予DRz164后小鼠未能形成对吗啡的位置偏爱。结论脱氧核酶DRz164在胞内能够切割Period1mRNA,抑制Period1基因表达,小鼠脑室注射DRz164可拮抗吗啡所引起的奖赏效应,缓解其对吗啡的精神依赖。Objective To investigate the regulatory effects of deoxyribozyme on the expression of Period 1 gene in vitro and on the morphine-induced psychic dependence in mice. Methods The specific deoxyribozymes toward Period 1 mRNA was designed by MFold analysis and synethsized chemically. By LipofectAMINE^TM mediated DNA transfection technique, DRz164 and pcDNA3-Perl were introduced into NIH3T3 cells. The effects of deoxyribozyme on Period 1 gene were studied by reverse transcript-polymerase chain reaction （RT-PCR） and flow cytometry（FCM）. The morphine-induced reward in mice was observed in a conditioned place preference test after pretreatment of the mice with the intracerebroventricular administration of deoxyribozyme. Results After NIH3T3 cells were transfected by pcDNA3-Perl and DRz164, the Period 1 mRNA was reduced by 42.4%. And PERIOD proteins were decreased by 57. 5%. After being pretreated with deoxyribozyme, the mice did not show morphine-induced place preference. Conclusion DRz164 can highly block the expression of Period 1 gene, which cleaves the Period 1 mRNA in the transfected cells specifically. The suppression of morphine-induced place preference can be effected by pretreating the mice with intracerebroventricular injection of deoxyribozyme for alleviating their psychic dependence on morphine.

关 键 词：PERIOD 1基因 脱氧核酶 基因表达 吗啡 条件位置偏爱 

分 类 号：R749.64[医药卫生—神经病学与精神病学]