单术者制作大鼠腹腔异位心脏移植模型操作技巧

Establishment skills of a rat model of abdominal heterotopic heart transplantation by single operator

目的探讨单术者建立大鼠腹腔异位心脏移植模型的制作流程及注意事项,为初学者提供详细的操作经验。方法选用雄性供体(Wistar)、受体(SD)大鼠68对,术前供、受体同时麻醉,受体备8/0线行腹主动脉、下腔静脉血管阻断,阻断背侧分支血管。供体左、右上腔静脉分别与肺门环扎,断升主动脉后灌注停跳液,游离肺动脉,穿横窦断肺动脉,冲洗心房,分离供体心脏。受体腹腔血管适当切口,6点与12点处褥式缝合,动脉连续缝合,静脉单针缝合,排气,再灌注。记录操作时间,行HE染色、检测CD_3、IL-1β及外周血CD_4^+T、CD_8^+T细胞。结果单术者行68例手术,手术时间(58.8±4.5)min,动、静脉吻合时间分别为(7.6±2.2)min和(13.5±4.2)min,供心总缺血时间(31.8±4.5)min,成功率为88.2%,术后第3、5天排斥反应较重,心肌组织间CD_3、IL-1β高表达,术后第1天起外周血CD_4^+T、CD_8^+T细胞逐渐增多。结论通过充分准备与熟练操作,单术者可实施成功率稳定的大鼠腹腔异位心脏移植模型。

Objective To explore the operation proeudure and notice of establishing a rat model of abdominal heterotopic heart transplantation by single operator, and summarize the detailed skill and experience for beginners. Methods 68 pairs male SD （recipients）/Wistar （donors） rats,preoperative anesthesia for recipients and donors at the same time,8/0 line was preparing for blocking recipients vena cava and abdominal aortic, blocking the branch vessel. Left and right superior vena ligature respectively with hilar, cutting the ascending aorta, perfusion cardioplegia, free puhnonary artery, across transverse sinus of pericardium for pulmonary artery cutting,atrial wash, free donor heart. Choosed appropriate incision of receptor abdominal blood vessels, mattress suture at 6 and 12 points, a single suture for vent gases,continuous suture in artery, single needle in vein, repeffusion after exhaust gas. Recording operation time, HE staining, IL-1β/CD3 immunohistoehemistry sliee, flow eytometry analysis of CD4^ ＋ T/CD8 ^＋ T lymphocytes subsets in the peripheral blood. Results Sixty-eight cases were treated by single operater, the operation time was （ 58.8 ±4.5 ） minutes, artery suture time was （7.6 ±2.2 ） minutes, vein suture time was （ 13.5 ± 4.2 ） minutes, the total donor heart isehemic time was（31.8 ±4.5）minutes,the success rate was 88.2%. The rejection reaction was stronger on the third and fifth day after surgery,with high expression of IL-1β/CD3 in cardiac allografts. The CD4^ ＋ T/CD8^ ＋ T lymphocytes subsets increased in the peripheral blood at first day after heart transplantation. Conclusion By fully preparation and skillfully operation, establishing a rat model of abdominal heterotopic heart transplantation by single operator has a stable success rate.