NO介导吗啡戒断大鼠脊髓Fos蛋白和NMDA_(1A)受体mRNA表达的增加

目的:观察NO在纳洛酮催促吗啡戒断大鼠脊髓神经元活动变化中的作用。方法:采用Fos免疫组织化学、NADPH-d组织化学、Fos/NADPH-d双标、鞘内注射、反义寡核苷酸和RT-RCR技术。结果:急性应用纳洛酮和慢性应用吗啡对大鼠脊髓Fos蛋白及NADPH-d阳性神经元表达无明显影响,二者也无Fos/NADPH-d双标神经元表达;纳洛酮催促吗啡戒断大鼠脊髓Fos蛋白、NADPH-d阳性神经元、纤维和终末表达明显增加,且出现Fos/NADPH-d双标神经元表达。预先鞘内注射nNOS反义寡核苷酸明显降低吗啡戒断症状评分,减少吗啡戒断大鼠脊髓Fos蛋白及NMDA_(1A)R mRNA表达。结论:NO介导吗啡戒断大鼠脊髓Fos和NMD_(1A)R mRNA表达的增加。

NO mediated increase of Fos protein and NMDA_(1A) R mRNA expression in rat spinal cord during morphine withdrawal

AIM: To investigate the effects of nitric oxide (NO) on activation of the rat spinal cord neurons during naloxone-precipitated morphine withdrawal. METHODS: Fos immunocytochemistry, NADPH-d histochemistry, Fos/ NADPH-d double-labeling, intrathecal injection, anti-sense oligonucleotides (AS-ONs) techniques, and RT-PCR were used. RESULTS: Acute administration of naloxone and chronic administration of morphine did not change the expression of Fos protein and NADPH-d positive neurons, and there was no expression of Fos/ NADPH-d double-labeled neurons in the spinal cord of rats. Morphine withdrawal increased the expression of Fos protein, NADPH-d positive, and Fos/NADPH-d double-labeled neurons, and they were observed in all the laminae of the rat spinal cord. Intrathecal injection of nNOS antisense oligonucleotides (nNOS-AS) inhibited the increase of Fos protein and NMDA1AR mRNA expression in the rat spinal cord during morphine withdrawal and decreased the scores of morphine withdrawal symptoms. The effect of nNOS-AS was greater than that of eNOS-AS. There was no effect in nNOS sense oligonucleotides (nNOS-S) group. CONCLUSION: NO mediated the increase of Fos protein and NMDA1AR mRNA expression in the rat spinal cord during morphine withdrawal.