小鼠腹部心脏移植模型的建立及技术改进

Establishment of abdominal cardiac transplant model in mice and technical modifications

背景:小鼠腹部心脏移植模型是研究免疫耐受的重要平台,虽然有各种改良方法的报道,但是最终均要行显微外科血管吻合。作者针对小鼠腹部心脏移植血管端侧吻合技术的一系列改良方法做了报道,改良的技术方法同样也适用于其他微血管端侧吻合。目的:建立稳定、可靠、易于掌握的小鼠腹部心脏移植模型。方法:实验动物供体为C57/BL6J(H-2b),DBA/2(H-2d),Balb/C(H-2d)小鼠共144只,每种各48只;受体为Balb/C(H-2d)共144只;由北京维通利华公司提供,实验方案经甘肃省人民医院动物实验伦理委员会批准(批准号为2017-011)。实验分为改良组和传统方法组,各施行心脏移植72对。改良组采用改良方法摘取供心;自制弧形血管夹阻断受体血管,11-0缝合针法切开血管前壁,170°双定点法吻合肺动脉与下腔静脉等方法施行小鼠腹部心脏移植;传统方法摘取供心采用游离升主动脉至头臂干处,游离肺动脉至左右分叉处,并分别离断的方法。比较2组血管吻合时间和移植成功率。结果与结论:①应用改良方法心脏移植成功率90.3%,供体手术时间(7.0±0.5)min,受体手术时间(53.5±5.9)min,血管阻断时间(30.4±4.5)min,静脉吻合时间(9.4±1.5)min;传统方法心脏移植成功率86.1%,供体手术时间(12.3±2.1) min,受体手术时间(80.0±7.1) min,血管阻断时间(45.6±6.0) min,静脉吻合时间(14.0±2.3) min,改良组心脏移植的时间及成功率显著优于传统方法组(P <0.01或P <0.05);②结果说明,改良方法提高了小鼠腹部心脏移植的速度,降低了手术难度,成功的建立了稳定,可靠的小鼠腹部心脏移植模型。

BACKGROUND: The mouse abdominal heart transplantation model is an important platform for studying immune tolerance. Although various modified methods have been reported, microsurgical vascular anastomosis is required. The authors have reported a series of modified methods for end-to-side anastomosis of mouse abdominal heart transplantation, and the modified technique is applicable to other microvascular end-to-side anastomosis.OBJECTIVE: To establish a stable, reliable and manageable abdominal heterotopic cardiac transplant model in mice.METHODS: One hundred and forty-four C57/BL6J(H-2b), DBA/2(H-2d), and Balb/C(H-2d) mice were as donors, n=48 per specie, and 144 Balb/C(H-2d) mice were as recipient mice. All mice were provided by Beijing Weitong Lihua Company, and the study was approved by the Experimental Animal Ethics Committee of Gansu Provincial Hospital(approval No. 2017-011). There were two groups, modified and traditional groups, and 72 pairs of mice underwent cardiac transplant. Abdominal heterotopic cardiac transplantation in mice was performed using the following techniques: donor heart was harvested by a modified approach;recipient blood vessels were blocked by a self-made arched vascular clamp;incision was made on the anterior wall of blood vessels using an 11-0 suture needle, and pulmonary artero-inferior vena cava anastomosis was established using 170-degree dual-fixation method. The venous anastomosis time and success rate of transplantation were compared.RESULTS AND CONCLUSION:(1) The success rate of transplantation in the modified group was 90.3%, the donor procedure time was(7.0±0.5) minutes, the recipient procedure time was(53.5±5.9) minutes, blood vessel blockage time was(30.4±4.5) minutes, and venous anastomosis time was(9.4±1.5) minutes. The success rate of transplantation in the traditional group was 86.1%;the donor procedure time was(12.3±2.1) minutes;the recipient procedure time was(80.0±7.1) minutes;blood vessel blockage time was(45.6±6.0) minutes;and venous anastomosis time was(14.0±2.3) minutes. The modified group was superior to the traditional group in the transplantation time and success rate(P < 0.01 or P < 0.05).(2) These results indicate that the modified approaches speed up the abdominal cardiac transplantation in mice and decrease the complexity of the surgery, and establish a stable and reliable abdominal heterotopic cardiac transplant model in mice.