B超引导下猕猴肝脏及肾脏组织的活体穿刺方法

Biopsy of liver and kidney tissues in rhesus monkeys under B-mode ultrasound guidance

目的建立B超引导下安全有效的猕猴活体穿刺以获取肝脏、肾脏样品的方法。方法 4只猕猴,雄性,8~12 kg,10~12岁,经5~10 mg/kg肌肉注射氯胺酮进行麻醉,麻醉后将其保定于手术台,侧仰卧位,于腹肋间区域、侧腰部肋缘下腹部区域剃毛及常规碘伏消毒。安装B超引导架于3.5 Hz彩色电脑声像仪探头上,于猕猴右上腹肋间区域、侧腰部肋缘下腹部区域进行探查寻找所要穿刺的肝脏或肾脏器官,My Lab 30CV B超进一步调节探头位置以便确认目标,注意避让门静脉及下腔静脉等大血管,将活检枪的射程调为15 mm,将穿刺针在穿刺引导架的引导下垂直经皮肤插入腹部约1 cm,观察B超实时影像,使穿刺目标与穿刺针位置在引导线内,即穿刺目标与穿刺针为垂直部位,按动穿刺枪扳机按钮进行穿刺,穿刺后迅速拔出穿刺针。将所获得的肝、肾组织进行RNA提取。结果每只动物每次穿刺均获取的肝或肾组织样品量经换算约13 mg,提取的RNA纯度高、完整性好,本法操作快速、可靠、安全,所有实验猴术后恢复良好,无大出血及死亡。结论本方法对建立超声引导下安全、有效获取恒河猴肝、肾组织样本,有着非常重要的意义。本方法能降低研究成本,并提高实验室非人灵长类动物的生活质量和动物福利,同时保证实验结果的准确性。

Objective To establish a safe and effective method of rhesus monkey biopsy to take liver and kidney samples under B-mode ultrasound guidance.Methods A total of 4 adult monkeys(weight:8-12 kg;sex:male;age:11-12 years old)were anaesthetized with 5-10 mg/kg of ketamine hydrochloride for each through intramuscular injection.After successful anesthesia,abdominal shaving and iodophor disinfection,they were monitored from intercostal area of right upper quadrant or lateral waist subcostal abdomen portions to find liver or kidney organ by MyLab 30CV B-mode ultrasonography with 3.5 Hz transducer which was fixed with a guiding frame.Large vessels such as the portal vein and inferior vena cava were carefully avoided.The range of the biopsy gun was set to 15 mm.When the puncture target and the puncture needle were positioned in the guide line,the puncture target was perpendicular to the puncture needle,and then the trigger button of the puncture needle was pressed to obtain the liver or kidney tissue samples respectively.After puncture,the needle was pulled out quickly.The obtained liver and kidney tissues were used to extract RNA.Results About 13 mg of liver or kidney tissue was obtained by each puncture with volume convertion.This method was fast,reliable and safe,and the total RNA had high purity and integrity.There was no postoperative bleeding and infection.Conclusions This is a very important method for obtaining liver and kidney tissue samples of rhesus monkeys with the guidance of ultrasound.With this method,the research cost can be reduced,the life quality and animal welfare of laboratory non-human primates can be improved,and the accuracy of experimental result can be ensured.