吗啡长时程作用对培养SK-N-SH细胞表达fosB基因的影响

Effects of long-term morphine on the expression of fosB gene in in vitro cultured SK-N-SH cells

目的探讨吗啡长时程作用时SK-N-SH细胞fosB基因表达的改变。方法建立慢性吗啡依赖SK-N-SH细胞模型,分别采用放射免疫法(radioimmunoassay,RIA)、电泳迁移率改变分析(electrophoresis mobility shift assay,EMSA)和RT-PCR检测cAMP(cyclic adenosine3′,5′-monophosphate)、CREB(cyclic AMP-responsive element-binding protein)的DNA结合活性和fosBmRNA表达。结果吗啡长时程作用SK-N-SH细胞CREB的DNA结合活性及fosBmRNA水平升高。结论吗啡长时程作用诱导SK-N-SH细胞fosB基因表达上调可能通过CREB的DNA结合活性升高而调控。

Objective To explore the effects of long-term morphine on the expression offosB gene in cultured SK-N-SH ceils. Methods Morphine-dependent SK-N-SH ceils were established by culturing SK-N-SH ceils with morphine for 48 h, then divided into morphine-dependent group and naloxone-withdrawn group in which the morphine-dependent SK-N-SH ceils were treated with 10μmol/L naloxone for 15 min. The SK-N-SH ceils cultured with physiological saline served as normal control group. The intracellular cAMP, the DNA-binding activity of CREB, and the expression offosB mRNA were detected by radioimmunoassay （RIA）, electrophoresis mobility shift assay （ EMSA）, and RT-PCR respectively. Results The intracellular cAMP, the DNAbinding activity of CREB, and fosB mRNA were significantly elevated in morphine-dependent and naloxonewithdrawn SK-N-SH ceils. Conclusion The upregulation offosB mRNA in SK-N-SH ceils treated with longterm morphine may be mediated through the activation of the DNA-binding activity of CREB.