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目的 观察吗啡依赖或吗啡戒断大鼠脊髓和脑干中一氧化氮合酶 (NOS)基因表达的变化。方法 以 β actin为内参照 ,用逆转录聚合酶链反应 (RT PCR)测定NOSmRNA的表达水平。结果 吗啡依赖大鼠脊髓和脑干NOS表达水平较正常对照大鼠降低 ,纳洛酮 ( 4mg·kg-1,ip)激发大鼠吗啡戒断症状 1h后脊髓和脑干中NOS表达水平明显升高 ,戒断 2h和 4h后NOS基因表达较 1h组减少。NOS抑制剂L N 硝基精氨酸甲酯 (L NAME ,10mg·kg-1)处理后大鼠吗啡戒断症状减少 ,同时脊髓和脑干的NOS基因表达水平较戒断 1h组明显降低。甲基东莨菪碱 ( 0 5mg·kg-1)处理组脊髓和脑干中NOS表达水平较戒断 1h组明显降低 ;选择性毒蕈碱受体M1拮抗剂 pirenzepine( 10mg·kg-1)处理组动物脊髓中NOS表达水平较戒断 1h组降低 ,而脑干中NOS表达水平没有改变 ;NMDA受体拮抗剂MK 80 1( 0 12 5mg·kg-1)处理后脊髓和脑干中NOS表达水平较戒断 1h组没有差异。结论 吗啡慢性处理后脊髓和脑干中NOSmR NA水平降低 ,抑制内源性NO生成和阻断毒蕈碱受体可以减少吗啡戒断所引起脊髓和脑干中NOS基因的表达
Objective To observe the changes of nitric oxide synthase (NOS) gene expression in spinal cord and brainstem of morphine dependent or morphine withdrawal rats. Methods β actin was used as an internal reference, and the expression of NOS mRNA was determined by reverse transcriptase-polymerase chain reaction (RT-PCR). Results NOS expression in the spinal cord and brainstem of morphine-dependent rats decreased compared with that of normal rats. Naloxone (4 mg · kg -1, ip) induced morphine withdrawal symptoms in rat spinal cord and brainstem after 1 h High, withdrawal 2h and 4h after NOS gene expression decreased compared with 1h group. The symptoms of morphine withdrawal in rats treated with NOS inhibitor L NAME (10 mg · kg-1) were decreased, and the expression of NOS gene in spinal cord and brainstem was significantly lower than that in the 1-h withdrawal group. The expression of NOS in spinal cord and brainstem of scopolamine treated group (0 5 mg · kg -1) was significantly lower than that of 1 h after abstinence treatment; the group of animals treated with selective muscarinic receptor antagonist pirenzepine (10 mg · kg -1) The expression of NOS in the spinal cord was lower than that in the abstinence group and the expression of NOS in the brainstem did not change. The expression of NOS in the spinal cord and brainstem after NMDA receptor antagonist MK 80 1 (0 12 5 mg · kg -1) Withdrawal 1h group no difference. Conclusions After chronic morphine treatment, the levels of NOSmRNA in the spinal cord and brainstem are decreased. Inhibition of endogenous nitric oxide production and blockade of muscarinic receptors may reduce the expression of NOS gene in spinal cord and brainstem caused by morphine withdrawal