Dalian Medical University, Dalian, Liaoning Province, China.
Dalian Medical University, Dalian, Liaoning Province, China.
Transplant Proc. 2021 Dec;53(10):3080-3086. doi: 10.1016/j.transproceed.2021.08.047. Epub 2021 Nov 5.
Renal transplantation is an effective treatment for end-stage renal disease, which involves pathophysiologic processes such as ischemia-reperfusion injury and immune rejection. The degree of ischemia-reperfusion injury is closely related to the functional state of the transplanted kidney. At present, the allogeneic kidney transplantation model has been widely used in related research. The traditional kidney transplantation model has the disadvantages of complicated vascular anastomosis, difficulty in ureteral reconstruction. The aim of this study was to establish a rat autologous orthotopic kidney transplantation model based on non-anastomotic technique.
Inbred Wistar rats weighing 260 to 280 g were selected. The rats were anesthetized by intraperitoneal injections of 40 mg/kg body weight pentobarbital sodium. We exposed and freed the left kidney after laparotomy and separated the left renal artery and left renal vein, abdominal aorta, and posterior vena cava. A purse-string suture with a diameter of 1 to 2 mm was made through the tunica media of the abdominal aorta. A puncture was made through the center of the purse-string suture. The in-dwelling needle was placed in the renal artery along the blood flow direction, and was infused with constant flow of 4°C heparinized lactated ringer's solution until the kidney became pale yellow. The renal vein was ligated and the renal artery was clamped. The in-dwelling needle was removed, purse-string suture was ligated, and the kidney was stored in a self-made autologous kidney transplant cold storage bag for 4 hours. We then opened the vein and artery, removed the cold storage bag, and rewarmed with 37°C normal saline. The abdomen was then closed layer by layer.
Fifty-two orthotopic renal transplantations were performed, which included pre-experimental (40 operations) and experimental stages (12 operations). The success rates of the 2 stages were 75% and 91.7%, respectively. The main causes of failure were intraoperative hemorrhagic shock and postoperative infection. The operation time of orthotopic renal transplantation was 360 ± 30 minutes, including 30 ± 10 minutes for dissociation and management of kidney and blood vessels, 1 ± 0.5 minutes for warm ischemia and 240 ± 10 minutes for cold storage. Rats were sacrificed at 1 day and 7 day respectively. The rats were in good condition after operation. They could eat and drink freely. At 24 hours and 1 week after transplantation, the kidney's blood supply was good, the intestine was light or showed no adhesions, and the abdominal cavity had no ascites or peculiar smell. Hematoxylin & eosin (H&E) staining showed that there were no obvious pathologic changes in the sham group. The orthotopic kidney transplantation 1-day group showed pathologic changes of ischemia-reperfusion, such as swelling, necrosis, shedding, and cast formation of renal tubular cells. The orthotopic kidney transplantation 7-day group recovered well, with mild dilation of the renal capsule and mild dilatation of the renal tubules.
The new model of autologous kidney transplantation is simple to use, does not require vascular anastomosis and ureteral reconstruction, and has a high success rate.
肾移植是治疗终末期肾病的有效方法，涉及缺血再灌注损伤和免疫排斥等病理生理过程。缺血再灌注损伤的程度与移植肾的功能状态密切相关。目前，同种异体肾移植模型已广泛应用于相关研究。传统的肾移植模型存在血管吻合复杂、输尿管重建困难等缺点。本研究旨在建立一种基于非吻合技术的大鼠自体原位肾移植模型。
选择体重 260-280g 的近交 Wistar 大鼠。大鼠经腹腔注射 40mg/kg 戊巴比妥钠麻醉后，行剖腹术暴露并游离左肾，分离左肾动脉、左肾静脉、腹主动脉和后腔静脉。用直径 1-2mm 的缝线在腹主动脉中膜做荷包缝合，在缝线中心做一个穿刺。将留置针沿血流方向插入肾动脉，用 4°C 肝素化乳酸林格氏液恒流灌注，直至肾脏呈淡黄色。结扎肾静脉，夹闭肾动脉。取出留置针，结扎荷包缝线，将肾脏放入自制自体肾移植冷藏袋中冷藏 4 小时。然后打开静脉和动脉，取出冷藏袋，用 37°C 生理盐水复温。然后逐层关闭腹部。
共进行 52 例原位肾移植术，包括预实验（40 例）和实验阶段（12 例）。2 个阶段的成功率分别为 75%和 91.7%。失败的主要原因是术中失血性休克和术后感染。原位肾移植的手术时间为 360±30 分钟，其中肾脏和血管的分离和处理时间为 30±10 分钟，热缺血时间为 1±0.5 分钟，冷保存时间为 240±10 分钟。大鼠分别在术后 1 天和 7 天处死。术后大鼠状况良好，饮食自由。移植后 24 小时和 1 周，肾脏血供良好，肠管轻或无粘连，腹腔无腹水或异味。苏木精和伊红（H&E）染色显示假手术组无明显病理改变。原位肾移植 1 天组表现为缺血再灌注损伤的病理变化，如肾小管细胞肿胀、坏死、脱落和铸型形成。原位肾移植 7 天组恢复良好，肾包膜轻度扩张，肾小管轻度扩张。
新的自体肾移植模型操作简单，无需血管吻合和输尿管重建，成功率高。
