一氧化氮合酶抑制剂抑制吗啡依赖及戒断大鼠脊髓神经元ERK的表达  

Nitric oxide synthase inhibitors suppression against expression of ERK in spinal cord of morphine dependent and withdrawal rats

作　　者：刘海林[1] 曹君利[2] 曾因明[2] 戴培静[1] 郑国龙[1] 

机构地区：[1]南京医科大学附属淮安市第一人民医院,江苏淮安223300 [2]江苏省麻醉学重点实验室,江苏徐州221002

出　　处：《中国药理学通报》2005年第11期1308-1312,共5页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No30200267);江苏省教委自然科学基金资助项目(No01KJD320036);江苏省淮安市2005年科技发展计划(NoHAS05029)

摘　　要：目的探讨鞘内分别注射非选择性一氧化氮合酶(NOS)抑制剂L-NAME、选择性神经元型一氧化氮合酶(nNOS)抑制剂7-硝基吲哚(7-NI)对吗啡戒断大鼠戒断症状和痛敏行为以及脊髓神经元p-ERK表达的影响。方法采用吗啡依赖及戒断模型,分为正常对照组、依赖组、戒断组、L-NAME组(L-NAME)、7-NI组(7-NI),分别作行为学评分(n=8)、免疫组织化学(n=6)和免疫印迹检测(n=4)。结果实验结果表明,①鞘内注射L-NAME、7-NI可明显减轻吗啡依赖大鼠戒断症状,戒断组戒断症状评分为28.6±4.89,L-NAME组、7-NI组分别为22.1±4.52(P<0.05)、16.2±3.99(P<0.01);戒断组促诱发痛评分(touch evoked agitationscores,TEA score)为13.5±2.55,L-NAME组、7-NI组分别为9.8±3.11(P<0.05)、7.5±2.56(P<0.01)。②鞘内注射L-NAME、7-NI可明显减少胸腰段脊髓背角Fos阳性神经元的数目,L-NAME组、7-NI组分别为293±47、267±52,均低于戒断组(380±71,P<0.05,P<0.01)。③L-NAME、7-NI组p-ERK阳性神经元的数目分别为46.8±11.58、40.5±8.55,均低于戒断组(66.6±11.6,P<0.05,P<0.01),两给药组脊髓p-ERK蛋白的表达也减少。④W estern b lot显示:鞘内注射NOS明显抑制吗啡戒断大鼠脊髓p-ERK蛋白表达增加。结论脊髓水平NO参与吗啡依赖和戒断反应,ERK信号通路可能介导NO的上述作用。Aim This study aimed to explore the effects of intrathecal injection of nitric oxide synthase inhibitors （L-NAME,7-NI） on morphine withdrawal response and the the spinal p-ERK expression. Methods Rats were divided into 5 groups： control group, dependence group, withdrawal group, L-NAME group（L- NAME, it） and 7-NI group （ 7-NI, it ）. Morphine withdrawal score, touch evoked agitation scores （TEA scores ）, immunohistochemical and western-blotting technique were used to evaluate morphine withdrawal response and the expression of Fos and p-ERK in the spinal cord, respectively. Results Intrathecal injection of non-specific NOS inhibitor L-NAME, nNOS inhibitor 7-NI significantly alleviated morphine withdrawal symptoms. Morphine withdrawal scores in the L- NAME（22. 1±4.52） and 7-NI groups（ 16. 2±3.99） were significantly lower than that of the withdrawal group（28.6±4.89） （P 〈O. 05,P 〈0.01）; TEA score of the withdrawal group was 13.5± 2.55, which was significantly higher than that of the L-NAME （9. 8±3. 11,P〈0.05） and 7-NI groups（7.5±2.56,P〈 0.01 ）. Fos-like positive neurons（Fos-LI） in the spinal dorsal horn of the withdrawal group were 380± 71, which were higher that of the L-NAME （293 ± 47, P 〈 0. 05） and 7-NI groups （228±49,P 〈0. 01）. Phospho-ERK positive neurons in the spinal dorsal horn of the L-NAME and 7-NI groups were 46. 8±11.58,40.5 ±8.55 respectively, which were significantly lower that of the withdrawal group （ 66.6 ± 11.6, P 〈 0. 05, P 〈0. 01 ）. Compared with the withdrawal group, level of p-ERK protein detected by Western blot in spinal cord of the L-NAME and 7-NI groups were significantly lower. Conclusion The spinal NO involves in morphine dependence and withdrawal, and its role may be mediated by ERK signal pathway.

关 键 词：吗啡依赖及戒断 脊髓 一氧化氮合酶 细胞外信号调节激酶 

分 类 号：R-332[医药卫生] R322.8