Animal Models Core

导演们从劳动密集度极高的创作任务 患有慢性疾病的动物，并照顾它们术后的即时恢复。核心还将提供 心力衰竭或假FIME发生前和发生期间的超声心动图评估 控制。在过去的10年里，动物模型核心在PPG中发挥了非常有效的作用。 各种动物模型将由库尔蒂斯·G·康尼什博士或在他的支持下制作，他已经 在慢性动物的准备方面有多年的经验，并且在这些特定的动物上有经验 准备工作。他将得到三名技术人员的协助，在动物使用前进行准备和护理 由PPG的投资人。该中心制作的模型将是：(1)心脏冠脉结扎模型 (2)兔心衰起搏模型。 所有动物的心脏状态的评估都是通过超声心动图获得的。核心是 由两个超声心动图支持。兔的成像是在Accuon Sequoia 5120机器上完成的， 啮齿动物的图像是在视觉超音速Vevo 770上获得的。约翰尼·哈克利女士主要负责 获取心力衰竭诱导前、中、后大鼠和兔的数据。 A.动物模型的外科准备 1.心衰大鼠模型：冠脉结扎。 SD大鼠(200+g，雄性)将被随机分为两组，HF组或Sham组 动物。心力衰竭动物的左冠状动脉将被从主动脉的分叉附近结扎。 假手术大鼠将接受同样的手术，但冠状动脉不会被结扎，这与 我们和其他实验室使用的程序。将大鼠用2%异氟醚麻醉，并给予 6pg阿托品/100克体重肌注。将经口插入气管插管，以便给药 正压通风。开胸手术将在左侧的4“^肋间间隙进行。 插管后，胸部将被刮毛，用外科肥皂擦洗，然后用倍他丁涂抹。全 在这个核心的手术将按照严格的无菌程序进行。心力衰竭组左冠状动脉 会用6-0Proline缝线结扎，正好在其从主动脉出口下方，在肺动脉流出之间 房束和左心房。在进行利多卡因之前，将注射0.5毫克的利多卡因，以帮助预防潜在的 致命的心律失常。充分的冠状动脉闭塞将由心脏增大和烫伤证明。 心肌。然后将胸管插入6‘*^肋间空间，并在此下方0.5厘米处出口 指向。在这些动作之后，胸腔将被关闭并被疏散。当老鼠开始呼吸时 胸腔内的液体和空气自然会被排出，胸管也会被拔掉。控制老鼠会 接受开胸手术和心脏手术，用利多卡因治疗，但冠脉结扎术不会 被安置好。在动物开始出现恢复迹象后，将拔掉气管插管，并 老鼠将被允许从麻醉中恢复。然后，这些老鼠将被放在单独的笼子里，直到 实验当天。 这些老鼠将被安置在一个有12小时明暗循环的房间里，环境温度保持在 22℃，湿度30-40%。实验室的食物(普利纳)和自来水将提供ACF锂。在我们的 脑梗塞大鼠的最终(立即或24小时)死亡率为21%。在完成后 手术大鼠将接受预防性的止痛药治疗：1.大约在恢复前45分钟 从麻醉开始，大鼠按0.05 mg/kg剂量给予丁丙诺啡； 丁丙诺啡0.05 mg/kg sq，每12小时1次。这将在手术后的头两天完成。老鼠会 在实验方案规定的时间内交付给PI。 234

Directors from the extremely labor intensive task of creating chronic animals and caring for their immediate post operative recovery. The Core will also provide echocardiographic assessment on animals prior to and during the development of heart failure or sham fime controls. The Animal Models Core has worked very effectively over the past 10 years of this PPG. Various animal models will be prepared by or under the supen/ision of Dr. Kurtis G. Cornish who has had many years of experience in the preparafion of chronic animals and is experienced in these specific animal preparations. He will be assisted by three technicians in the preparation and care of animals prior to their use by the PPG invesfigators. The models prepared by this Core will be (1) the coronary ligafion model of heart failure (HF) in the rat, and (2) the pacing model of HF in the rabbit. Assessment of the cardiac status of all animals is obtained echocardiographically. The Core is supported by two echocardiographs. Rabbit imaging is done on an Accuson Sequoia 5120 machine and rodent images are obtained on a Visual Sonics Vevo 770. Ms. Johnnie Hackley is primarily responsible for obtaining the data on the rats and rabbits prior to, during and after the inducfion of heart failure. A. Surgical Preparation of Animal Models 1. Rat model of heart failure: coronary ligation. Sprague-Dawley rats (200+ g, males) will be assigned randomly to one of two groups, HF or sham animals. The heart failure animals will have their left coronary artery ligated near its bifurcafion from the aorta. Sham rats will undergo the same surgery but the coronary arteries will not be tied, consistent with the procedure used by us and by other laboratories^'^¿. The rat will be anesthefized with 2% Isoflurane and given 6 pg atropine/100 gm body weight im. An endotracheal tube will be inserted orally in order to administer positive pressure ventilation. A thoracotomy will be done at the 4"^ intercostal space on the left side. Once intubated, the chest will be shaved and scrubbed with surgical soap and then painted with Betadine. All surgeries in this Core will be done following strict sterile procedures. In the HF group, the left coronary artery will be ligated with 6-0 Proline suture, just below its exit from the aorta, between the pulmonary artery outflow tract and left atrium. Prior to the ligafion, 0.5 mg of lidocaine will be injected SQ to help prevent potenfially lethal arrhythmias. Adequate coronary occlusion will be evidenced by cardiac enlargement and blanching of the myocardium. A chest tube will then be inserted in the 6'*^ intercostal space and exited 0.5 cm below this point. Following these maneuvers the thorax will be closed and evacuated. As the rats begin to breathe spontaneously, the fluid and air in the chest will be evacuated and the chest tube removed. Control rats will undergo thoracotomy and the manipulafion of the heart, treated with lidocaine, but the coronary ligature will not be placed. After the animal begins to show signs of recovery, the endotracheal tube will be removed and the rats will be allowed to recover from the anesthesia. The rats will then be placed in individual cages until the day of the experiment. The rats will be housed in a room with a 12-hour light-dark cycle, an ambient temperature maintained at 22¿C and humidity of 30-40%. Laboratory chow (Purina) and tap water will be available acf libitum. In our hands there is an inifial (immediately or 24 hour) mortality rate of 21% for the infarcted rats. After completion of surgery the rats will be treated prophylactically with analgesics: 1. approximately 45 minutes before recovery from the anesthetic the rats will be given Buprinorphine at a dose of 0.05 mg/kg; 2. Then they will receive Buprinorphine 0.05 mg/kg SQ every 12 hours. This will be done for the first 2 post-operative days. Rats will be delivered to the PI at the time prescribed by the experimental protocol. 234