GLUT1在甲基苯丙胺与HIV-Tat蛋白协同诱导大鼠血脑屏障通透性改变中的作用研究

Role of GLUT1 in Synergistic Induction of Blood-brain Barrier Permeability in Rats by Methamphetamine and HIV-Tat Protein

目的观察甲基苯丙胺(METH)与HIV-Tat蛋白协同诱导大鼠血脑屏障通透性的改变,探讨GLUT1在METH与HIV-Tat蛋白协同诱导血脑屏障通透性改变中的作用。方法40只雄性SD大鼠随机分成4组,对照组(每日10mL·kg^-1生理盐水)、METH组、HIV-Tat蛋白组、METH+HIV-Tat蛋白组。建立血脑屏障损伤的大鼠模型:采用腹腔注射METH(每日10mg·kg^-1)和尾静脉注射HIV-Tat蛋白(每日400ng·kg^-1)的方法。模型建成功后检测各组脑区伊文思蓝(EB)的含量,并通过Western-Blotting和免疫组织化学观察各组脑中GLUT1表达情况。结果与对照组相比,METH组、HIV-Tat蛋白组和METH+HIV-Tat蛋白组脑组织的EB含量明显增加;分别与METH组和HIV-Tat组相比,METH+HIV-Tat蛋白组脑组织内EB含量明显升高。此外,免疫组化和WB结果显示,与对照组相比,METH组、HIV-Tat蛋白组和METH+HIV-Tat蛋白组表达GLUT1的水平不同程度降低(P<0.01);分别与METH组和HIV-Tat蛋白组相比,METH+HIV-Tat蛋白组GLUT1的表达水平明显降低(P<0.05)。结论METH和HIV-Tat蛋白可协同作用诱导大鼠血脑屏障通透性发生改变,而GLUT1在METH和HIV-Tat蛋白的协同作用中可起调节作用。

Objective To investigate the synergistic effect of methamphetamine (METH) and HIV-Tat protein on the permeability of blood-brain barrier in rats,and to explore the role of GLUT1 during the course. Methods Forty male Sprague-Dawley rats were randomly divided into 4 groups:control group (10 mL·kg^-1 NS per day),METH group,HIV-Tat protein group and METH+HIV-Tat protein group.A rat model of blood-brain barrier injury was established by intraperitoneal injection METH (10 mL·kg^-1 per day) and tail vein injection HIV-Tat protein (400 ng·kg^-1 per day).After the model was established,the content of Evans blue (EB) in each group was detected,and the expression levels of GLUT1 were observed by Immunohistochemistry and Western-Blotting. Results Compared with the control group,the EB content of the METH group,the HIV-Tat protein group and the METH+HIV-Tat protein group were significantly increased.Compared with METH group and HIV-Tat group,respectively,the EB content of METH+HIV-Tat group was significantly increased.In addition,the immunohistochemistry and WB results showed that the expression levels of GLUT1 were decreased to some extent in the METH group,HIV-Tat protein group and METH+HIV-Tat protein group,compared with the control group ( P <0.01).The expression level of GLUT1 in the METH+HIV-Tat protein group was significantly decreased compared with the METH and HIV-Tat protein group,respectively ( P <0.05). Conclusion METH and HIV-Tat proteins could synergistically alter the blood-brain barrier permeability,and GLUT1 involved in the regulation of synergistic effects of METH and HIV-Tat proteins.