Effects of direct transduction of neuronal nitric oxide synthase protein into the nucleus tractus solitarius on circulation

背景：孤束核(NTS)是外周压力感受器初级传入纤维的主要终末部位。有报道称双侧微量注射一种反义神经元型一氧化氮合酶(NNOS)。进入NTS的寡核苷酸可增加动脉血压(ABP)。在以前的研究中，我们证明了通过微量注射日本包膜血凝病毒(HvJ-E)载体的β-半乳糖苷酶(β-GAL)，将蛋白质直接转导到大鼠特定的限制性脑区是可能的。然而，目前还不知道转导的蛋白质是否完成了它的功能。方法：雄性Wistar大鼠，体重250~350g，用乌拉坦(1.45g/kg，ip)麻醉。连续测量平均动脉压(ABP)、平均动脉压(MABP)和心率(HR)。双侧…显微注射携带hvj-E载体的一氧化氮合酶更多的是在NTS。作为对照，将β-Gal蛋白与hvj-E载体进行同样的显微注射。实验结束后取出延髓，进行免疫组织化学染色。结果：NTS内微量注射nNOS后1h-3h，mABP较对照组明显降低。(1H：nNOS，OH值的94%，n=6；3-Gal，106%，n=9；p&lt；0.05。2H：nNOS，89%；/3-Gal，106%，n=9；p&lt；0.01。结果：NTS内微量注射nNOS后1h-3h，mABP明显低于对照组。(1H：nNOS，OH值的94%，n=6；3-Gal，106%，n=9；p&lt；0.05。2H：nNOS，89%；/3-Gal，106%，n=9；p&lt；0.01。结果：将nNOS与HVJ-E载体一起微量注射至NTS，1h后mABP较对照组显著降低。(1H：nNOS，OH值的94%，n=6；3-Gal，106%，n=9；p&lt；0.05。2H：nNOS，89%；/3-Gal，106%，n=9；p&lt；0.01。~H：nNOS，86%；3-Gal，106%，n=9；p&lt；0.01。我们认为，我们在这里提出的类型的靶向递送系统在向中枢神经系统给药治疗性蛋白质方面可能有广泛的应用。较少

BACKGROUND: The nucleus tractus solitarii (NTS) is known to be the principal termination site of the primary afferent fibers from peripheral barorecepters. It has been reported that bilateral microinjection of an antisense neuronal nitric oxide synthase (nNOS). oligonucleotide into the NTS increases in arterial blood pressure (ABP). In the previous study, we demonstrated that direct in vivo protein transduction into a specific restricted brain area is possible in rats by microinjecting β-galactosidase (β-gal) with hemagglutinnating virus of Japan envelope (HVJ-E) vector. However, it is not known whether the transducted protein fulfills its function. In the present study, we examined the effect of direct in vivo nNOS protein transduction into the NTS on circulation.METHODS: Male Wistar rats, weighing 250-350 g, were anesthetized with urethane (1.45 g/kg, i.p.). The ABP, mean ABP (mABP), and heart rate (HR) was measured continuously. The nNOS with HVJ-E vector was microinjected bilateral … More ly into the NTS. As a control β-gal protein with HVJ-E vector was microinjected similarly. After the experiment, the medulla oblongata was removed and the immunohistochemical staining was performed.RESULTS: Microinjection of nNOS with HVJ-E vector into the NTS produced a significant decrease in mABP compared with the control at 1h - 3h. (1h: nNOS, 94% of the value at Oh, n=6; 3 -gal, 106%, n=9; p<0.05. 2h: nNOS, 89%; /3 -gal, 106%, n=9; p<0.01. 3h: nNOS, 86%; 3 -gal, 106%, n=9; p<0.01.)RESULTS: Microinjection of nNOS with HVJ-E vector into the NTS produced a significant decrease in mABP compared with the control at 1h - 3h. (1h: nNOS, 94% of the value at Oh, n=6; 3 -gal, 106%, n=9; p<0.05. 2h: nNOS, 89%; /3 -gal, 106%, n=9; p<0.01. 3h: nNOS, 86%; 3 -gal, 106%, n=9; p<0.01.)RESULTS : Microinjection of nNOS with HVJ-E vector into the NTS produced a significant decrease in mABP compared with the control at lh - 3h. (1h:nNOS, 94% of the value at Oh, n=6 ; 3 -gal, 106%, n=9 ; p<0.05. 2h:nNOS, 89% ; /3 -gal, 106%, n=9 ; p<0.01. 3h:nNOS, 86% ; 3 -gal, 106%, n=9 ; p<0.01.)CONCLUSION : We demonstrated here that the transducted protein fulfills its function. We suggested that the type of targeted delivery system we present here may have wide applications in the administration of therapeutic proteins to the central nervous system. Less