Development of an in vivo real-time measurement system of nitric oxide and superoxide and their clinical applications

导管型NO传感器的研制：首先，我们评估了体内NO传感器的基本性能。在NO气体饱和纯水中，该传感器的灵敏度约为580pA/nM-NO。该传感器对NO具有较高的选择性。静脉注射硝酸甘油后，麻醉大鼠腹主动脉血中NO含量升高至NM水平。其次，通过将体内NO传感器包裹在4F导管中，研制了一种原型导管型NO传感器。将该传感器插入麻醉犬的胸主动脉。静脉注射乙酰胆碱(ACh)后，血浆NO水平一过性升高。L-尼莫地平抑制NO合成后，ACh引起的血浆NO水平升高明显低于L-尼莫地平治疗前。最后，我们研制了一种用于近端血管NO测量的导管式NO传感器。一个柔软的尖端固定在检测尖端的边缘。该传感器被插入麻醉犬的冠状静脉窦内。冠脉内注射ACh可使血浆NO水平升高至5 nM。总之，新研制的导管型NO传感器具有良好的性能，可用于临床评价内皮功能。超氧化物传感器的研制：通过化学修饰将超氧化物歧化酶固定在铂丝上，制备了超氧化物歧化酶。该传感器安装在流体式测量系统中。将分离的器官(心脏、肝脏和肾脏)加载到系统中，可以成功地测量这些器官释放的超氧阴离子水平。

Development of a catheter-type NO sensor : First, we evaluated the basic performance of an in vivo NO sensor. Using a NO gas-saturated pure water, the sensor showed a sensitivity of about 580 pA/nM-NO. The sensor also demonstrated high selectivity to NO. When nitroglycerin was infused, plasma NO level in the abdominal aorta of an anesthetized rat increased to some nM level. Secondly, a proto-type catheter-type NO sensor was developed by encasing the in vivo NO sensor in a 4F catheter. The sensor was inserted into the thoracic aorta of an anesthetized dog. Plasma NO level increased transiently by acetylcholine (ACh) infusion. After inhibition of NO synthesis by L-NMMA, the ACh-induced increase in plasma NO level was significantly lower than that before L-NMMA. Finally, we have developed a catheter-type NO sensor for NO measurement in the proximal vessels. A soft tip was fixed at the edge of the detection tip. The sensor was inserted into the coronary sinus of an anesthetized dog. Intra-coronary infusion of ACh increased plasma NO level up to 5 nM. In conclusion, the newly developed catheter-type NO sensors demonstrated satisfying performance and would be applicable to clinical evaluation of endothelial function.Development of a superoxide sensor : A superoxide was prepared by fixing SOD on a Pt wire with some chemical modification. The sensor was located in a flow-cell-type measurement system. Isolated organs (heart, liver, and kidney) were loaded into the system, and superoxide level released from these organs could be measured successfully.