【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。更多还原

【Abstract】 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-Per1 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-Per1 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.更多还原