吗啡耐受过程中Let-7c和μ型阿片受体的作用研究

The function analysis for Let-7c and μ opioid receptor in morphine tolerance

目的探讨吗啡耐受过程中Let-7c和μ型阿片受体(MOR)的具体变化及Let-7c对MOR影响的研究。方法通过吗啡体外刺激SH-SY5Y细胞后提取其总蛋白和总RNA,Western Blot检测MOR的改变情况,RT-qPCR方法检测该过程中Let-7c的改变情况,分析MOR在吗啡和Let-7抑制剂(Anti-Let-7c)联合作用下的变化情况,并使用双荧光素酶报告系统对Let-7c与MOR结合位点进行验证。结果吗啡体外作用SH-SY5Y细胞后,MOR表达降低,72h降低较明显,同时Let-7c表达量进行性升高(P<0.01)。在Anti-Let-7c和吗啡联合作用下,吗啡降低MOR的作用减弱(P<0.01),Let-7c与MOR存在结合位点。结论吗啡能降低MOR在SH-SY5Y细胞的表达,Let-7c为吗啡降低MOR表达的上游分子信号,抑制Let-7c后能减少MOR丢失,从而提高镇痛疗效。

Objective To investigate the gene expression profile of morphine tolerance and correlation between Let-7candμopioid receptor（MOR）in this process.Methods SH-SY5 Ycells were stimulated by morphine for 24hand/or 48 h,and then the total proteins and RNAs were extracted,respectively.The expression of MOR and the changes of Let-7cwas detected by Western blot and RT-PCR respectivelyl.MOR in SH-SY5 Ycells were treated in the combination of morphine and Let-7cinhibitor.Results In the SH-SY5 Ycells,treated by morphine for 48 hor 72h,the expression level of MOR was reduced,and the expression of 72 hlower than 48 h.Interesting,the reduction of MOR by morphine was weaken in the combination with Let-7cinhibitor.Conclusion The expression of MOR in SH-SY5 Ycells were treated by morphine was reduced,and Let-7cfunction as an upstream molecule in this process.Our results indicate that Let-7cmay participate in the regulatory mechanisms of MOR associated with the pain.