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

背景技术背景：孤束核（NTS）是外周压力感受器初级传入纤维的主要终止部位。已有报道双侧显微注射反义神经元型一氧化氮合酶（nNOS）。将寡核苷酸导入NTS会增加动脉血压（ABP）。在先前的研究中，我们证明了在大鼠体内直接将蛋白质转导到特定的限制性脑区是可能的，通过显微注射β-半乳糖苷酶（β-gal）与日本血凝病毒包膜（HVJ-E）载体。然而，尚不清楚转导的蛋白质是否实现其功能。在本研究中，我们研究了直接在体内的nNOS蛋白转导到NTS上circulation.METHODS的效果：雄性Wistar大鼠，体重250-350 g，麻醉与乌拉坦（1.45 g/kg，i. p.）。连续测量ABP、平均ABP（mABP）和心率（HR）。将携带HVJ-E载体的nNOS双侧显微注射 关于我们 进入NTS。作为对照，类似地显微注射具有HVJ-E载体的β-gal蛋白。结果：NTS内微量注射nNOS和HVJ-E载体后，mABP在1 ~ 3 h较对照组明显降低。(1h：nNOS，0 h时值的94%，n=6; 3 -gal，106%，n=9; p<0.05。2h：nNOS，89%; β-gal，106%，n=9; p<0.01。3h：nNOS，86%; 3 -gal，106%，n=9; p<0.01。）结果：NTS内微量注射nNOS和HVJ-E载体后，mABP在1h ~ 3 h较对照组明显降低。(1h：nNOS，0 h时值的94%，n=6; 3 -gal，106%，n=9; p<0.05。2h：nNOS，89%; β-gal，106%，n=9; p<0.01。3h：nNOS，86%; 3 -gal，106%，n=9; p<0.01。）结果：NTS内微量注射nNOS和HVJ-E载体后，mABP在1h-3 h较对照组明显降低。(1h：nNOS，0 h时值的94%，n=6 ; 3 -gal，106%，n=9 ; p<0.05。2h：nNOS，89% ; β-gal，106%，n=9 ; p<0.01。3h：nNOS，86% ; 3 -gal，106%，n=9 ; p<0.01。）结论：我们证明了转导蛋白的功能。我们建议，我们在这里提出的靶向递送系统的类型可能有广泛的应用在中枢神经系统的治疗性蛋白质的管理。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