细胞水平吗啡耐受相关基因的分离

In the present study, we found the following results: 1.a novel human gene MADP-1 related to morphine tolerance has been isolated, cloned and chromosomal localized in SH-SY5Y cell line using differential display PCR.It was found that this gene upregulated after morphine treatment in SH-SY5Y cell line confirmed by Northern Blotting analysis.MADP is a RNA binding protein,encoding 217 amino acids, has binding sites of several transcriptional factors in the upstream of the coding area suggest that it might be regulated by several transcriptional factors. In order to further study the function of this gene, we got the in vivo and in vitro expression vectors of this gene. .2.72 differential genes after morphine treatment were found as compared to control group in SH-SY5Y cell line with DNA microarray analysis, 52 of them upregulated and 20 down-regulated after morphine tolerance. .3. we also found that the gene expression level of cyclin-dependent kinase 5 (CDK5), a cell-cycle related kinase，increased at 8h and peaked at 72h after morphine treatment. Protein expression level of this gene showed similar pattern suggest that this gene may relate to morphine tolerance. .To elucidate the function of these genes helps to understand the neurobiological mechanisms of morphine tolerance and addiction.

吗啡耐受现象具有复杂的神经生物学机制。本课题采用反相分子生物学最新发展起来的ｍＲＮＡ差别显示技术，分离对吗啡产生耐受的细胞在表达水平出现差异的基因，即吗啡耐受的相关基因。从分子水平研究相关基因及其编码的新型多肽，及其在吗啡耐受现象中的功能作用。本研究可能为防止阿片耐受的形成提供理论依据。