Hirooka Y, Sakai K, Kishi T, Takeshita A
Department of Cardiovascular Medicine, Cardiovascular Science, Graduate School of Medical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan.
Ann N Y Acad Sci. 2001 Jun;940:197-205. doi: 10.1111/j.1749-6632.2001.tb03677.x.
The nucleus tractus solitarii (NTS) is an important site for the regulation of sympathetic nerve activity. It receives the signals through afferent fibers from arterial baroreceptors, chemoreceptors, cardiopulmonary receptors, and other visceral receptors. Many studies have examined the role of nitric oxide (NO) in the NTS in cardiovascular regulation. However, most of these studies were conducted in an acute state with anesthesia. We have developed a novel technique of endothelial nitric oxide synthase (eNOS) gene transfer into the NTS in vivo. Adenovirus vectors encoding either the beta-galactosidase gene (Ad beta gal) or the endothelial nitric oxide synthase gene (AdeNOS) gene were transfected into the NTS. In the Ad beta gal-treated rats, the local expression of beta-galactosidase was confirmed by X-Gal staining, and beta-galactosidase activity was quantified using a colorimetric assay. In the AdeNOS-treated rats, the local expression of eNOS protein was confirmed by immunohistochemistry, and eNOS production was measured by in vivo microdialysis. Blood pressure and heart rate were monitored by a radiotelemetry system in a conscious state. The expression of each gene was observed from day 5 to day 10 after the gene transfer. In the AdeNOS-treated rats, blood pressure and heart rate significantly decreased from day 5 to day 10, and then thereafter gradually recovered over time. Our method may be useful in examining the local effect of a particular substance produced by a specific gene in the brain on cardiovascular function.
孤束核（NTS）是调节交感神经活动的重要部位。它通过来自动脉压力感受器、化学感受器、心肺感受器及其他内脏感受器的传入纤维接收信号。许多研究探讨了一氧化氮（NO）在NTS对心血管调节中的作用。然而，这些研究大多是在麻醉的急性状态下进行的。我们开发了一种将内皮型一氧化氮合酶（eNOS）基因体内转移至NTS的新技术。将编码β-半乳糖苷酶基因（Adβgal）或内皮型一氧化氮合酶基因（AdeNOS）的腺病毒载体转染至NTS。在Adβgal处理的大鼠中，通过X-Gal染色确认β-半乳糖苷酶的局部表达，并使用比色法对β-半乳糖苷酶活性进行定量。在AdeNOS处理的大鼠中，通过免疫组织化学确认eNOS蛋白的局部表达，并通过体内微透析测量eNOS的产生。在清醒状态下通过无线电遥测系统监测血压和心率。在基因转移后第5天至第10天观察每个基因的表达。在AdeNOS处理的大鼠中，血压和心率从第5天至第10天显著下降，此后随时间逐渐恢复。我们的方法可能有助于研究大脑中特定基因产生的特定物质对心血管功能的局部影响。
