论文部分内容阅读
目的观察甲基苯丙胺(MA)与HIV-Tat蛋白协同作用致大鼠相关脑区N-甲基-D-天冬氨酸(NMDA)受体NR2B亚基和诱导型一氧化氮合成酶(iNOS)的表达变化,探讨MA与HIV-Tat协同作用对神经系统的影响.方法 50只健康雄性SD大鼠随机分为实验组:MA组(10 mg/kg MA,每日2次腹腔注射,连续4 d)、HIV-Tat组(10μg HIV-Tat注入大鼠脑纹状体)和MA+HIV-Tat组(按MA组注射MA 4 d后按HIV-Tat组注入HIV-Tat);对照组:生理盐水腹腔注射或纹状体内注射.各组大鼠分别于注射结束后48 h和7 d处死,麻醉后用多聚甲醛灌注,取出脑组织并固定,石蜡包埋,作冠状切面,用免疫组化和蛋白免疫印迹法对中毒大鼠纹状体中NMDA受体NR2B亚基和iNOS的表达进行观察,并进行图像分析,测量其平均光密度值,与对照组比较并进行统计学分析.结果各实验组与对照组比较,纹状体内NMDA受体NR2B亚基和iNOS的表达均有不同程度增高,差异具有统计学意义(P<0.05);其中MA+HIV-Tat组与MA组、HIV-Tat组比较,NMDA受体NR2B亚基和iNOS的表达显著增高(P<0.05).结论 MA与HIV-Tat蛋白的共同作用能够显著增加NMDA受体NR2B亚基和iNOS的表达,揭示NMDA受体NR2B亚基和iNOS参与了MA与HIV-Tat蛋白的协同神经毒性机制.
Objective To observe the synergistic effect of methamphetamine (MA) and HIV-Tat protein on N-methyl-D-aspartate (NMDA) receptor NR2B subunit and inducible nitric oxide synthase (iNOS ) To investigate the synergistic effects of MA and HIV-Tat on the nervous system.Methods Fifty healthy male Sprague-Dawley rats were randomly divided into experimental group (MA group, 10 mg / kg MA, twice daily ip, continuous 4 d), HIV-Tat group (10 μg HIV-Tat injected into rat brain striatum) and MA + HIV-Tat group (injected with HIV-Tat by HIV-Tat group MA 4 days after MA injection) : Intraperitoneal injection of saline or intracerebral injection.The rats in each group were sacrificed at 48 h and 7 d after the injection, and were perfused with paraformaldehyde after anesthesia, the brain tissue was removed and fixed, embedded in paraffin, coronal section Immunohistochemistry and Western blotting were used to detect the expression of NMDA receptor NR2B subunit and iNOS in the striatum of striatum, and the images were analyzed. The average optical density was measured and compared with the control group for statistical analysis Results Compared with the control group, the expression of NMDA receptor NR2B subunit and iNOS in striatum were increased in all groups, the difference was statistically significant ( P <0.05). The expression of NMDA receptor NR2B subunit and iNOS in MA + HIV-Tat group was significantly higher than that in MA group and HIV-Tat group (P <0.05) .Conclusion The interaction between MA and HIV-Tat protein Can significantly increase the expression of NMDA receptor NR2B subunit and iNOS, revealing NMDA receptor NR2B subunit and iNOS involved in the synergistic neurotoxic mechanism of MA and HIV-Tat protein.