吗啡对原代培养大鼠海马神经元CCK受体mRNA表达的影响

目的探讨原代培养大鼠海马神经元上胆囊收缩素(cholecystokinin,CCK)受体CCK-AR及CCK-BR mRNA表达及吗啡对其表达的影响。方法采用新生大鼠海马神经元无血清原代培养技术,用吗啡(100μmol·L-1培养基)孵育3,6,12,18,24,36,48,72h,以及采用不同浓度吗啡(10,100,150μmol·L-1)孵育6、30h后,RT-PCR及测序方法观察CCK-AR及CCK-BR mRNA表达及吗啡对其表达的影响。结果RT-PCR结果显示,CCK-AR和CCK-BR mRNA在原代大鼠海马神经元上均有表达;CCK-AR mRNA RT-PCR扩增产物为218bp,CCK-BR mRNA RT-PCR扩增产物为444bp,经测序证实结果的可靠性;CCK-BR mRNA的表达量明显高于CCK-AR。吗啡作用后可使2种CCK受体mRNA表达上调。吗啡浓度及作用时间的不同对CCK-AR和CCK-BRmRNA表达的影响不同。结论原代大鼠海马神经元上有CCK-AR和CCK-BR,以CCK-BR为主;吗啡可使2种CCK受体mRNA表达上调。

Effect of expression of cholecystokinin receptor mRNA in primary culturing hippocampus neurons of newborn rat induced by morphine

Aim To investigate the expression of cholecystokinin (CCK) receptor mRNA and the effect of morphine on its expression in primary culturing hippocampus neurons of newborn rat. Methods By means of the technique of primary culturing hippocampus neurons of newborn rat, the primary hippocampus neurons were cultured with serum-free medium, and incubated with different dosage of morphine (10,100,150μmol·L~ -1 ) for 6 and 30 h, and incubated with morphine (100 μmol·L~ -1 ) for 3, 6,12,18,24,36,48,72 h respectively. The expression of CCK-AR and CCK-BR and effects of morphine on their expression were investigated by RT-PCR in primary culturing hippocampus neurons of newborn rat. Results The amplification products of CCK-AR and CCK-BR were approximately 218 bp and 444 bp respectively, which were proved by sequencing. The expression level of CCK-BR mRNA was higher than that of CCK-AR mRNA. The exprssion of CCK-AR and CCK-BR mRNA were obviously up-regulated by morphine. The expression of CCK-AR and CCK-BR mRNA were differently regulated by different dosage of morphine or different incubation time.Conclusion Both CCK-AR and CCK-BR mRNA are expressed in primary culturing hippocampus neurons of newborn rat and the major type is CCK-BR. The expression of two types of CCK receptor can be up-regulated by morphine.