[Inhibitory effects of deoxyribozyme on the expression of Period1 gene in vitro and on morphine-induced psychic dependence in mice].
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 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.


CONCLUSIONS
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 alleviating their psychic dependence on morphine.
Zhou
Wei
W
Department of Biomedical Engineering, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China.
Liu
Yan-you
YY
Wang
Yue-qi
YQ
Wang
Yu-hui
YH
Xiao
Jing
J
Zhu
Bin
B
Peng
Wen-zhen
WZ
Wang
Zheng-rong
ZR
D000818:Animals; D021881:DNA, Catalytic / Q000494:pharmacology*; D005136:Eye Proteins / Q000096:biosynthesis* / Q000235:genetics; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D009021:Morphine Dependence / Q000235:genetics / Q000378:metabolism* / Q000523:psychology; D056950:Period Circadian Proteins; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D011897:Random Allocation
D021881:DNA, Catalytic; D005136:Eye Proteins; C539023:Per1 protein, mouse; D056950:Period Circadian Proteins; D012333:RNA, Messenger