Beagle犬自体肾移植模型建立

The establishment of Beagle dog autologous kidney transplantation model

目的完善Beagle犬自体肾移植模型构建方法,总结成功建立模型的经验。方法选取18只成年Beagle犬随机分为假手术组和模型组,每组各9只。假手术组保留左肾,摘除右肾;模型组采用自体左肾异位肾移植,摘除右肾,实验操作者均为同一组医师。分布于术后第1、2、3、5、7、10、13、16、19、22、25和28天上午8点抽取静脉血,检测血清肌酐和尿素氮。采用两独立样本t检验比较假手术和模型组Beagle犬术后28天内血清肌酐和尿素氮。P<0.05为差异有统计学意义。结果模型组1只实验犬因血清肌酐持续升高且无好转迹象,于术后第7天行安乐死,解剖发现肾动脉血栓形成,移植肾坏死。其余8只模型组实验犬血清肌酐和尿素氮术后第2天短暂升高后逐渐降低并保存稳定,术后第1、2、3、5、7、10、13、16和19天血清肌酐和尿素氮水平均高于假手术组,差异均有统计学意义(t=-4.544、-5.083、-5.171、-4.946、-5.460、-5.345、-3.757、-3.918和-3.435;t=-4.810、-8.119、-10.379、-12.789、-15.210、-9.983、-5.014、-4.599和-2.657,P均<0.05)。模型组观察28天后予安乐死处理,解剖可见移植肾大小约7 cm×4 cm×2 cm,质地适中,肾周有肠道包裹,肾动脉与髂内动脉无血栓形成,肾静脉与髂外静脉回流正常。重建血管血流情况良好,输尿管及膀胱吻合口处未见狭窄。假手术组解剖后未见任何异常,剩余左肾呈现正常解剖形态,肾动脉、肾静脉与输尿管形态结构、位置关系均表现为正常生理状态。模型组实验犬移植肾组织HE染色光镜下可见肾组织结构改变轻微,肾小球分布均匀一致,包曼氏囊未见异常改变;肾小管上皮细胞呈方形或柱状,胞浆丰富,管腔结构完整,未见明显损伤;肾盂内可见中性粒细胞及浆细胞浸润。结论通过实验成功摸索出Beagle犬自体肾移植围手术期管理方法,可使该动物模型制作简便、稳定性高,具有可重复性。

Objective To improve the method of autologous kidney transplantation model construction in Beagle dogs and to summarize the experience of successful establishment of the model.Methods Eighteen adult Beagle dogs were randomly divided into sham operation group and model group,with 9 dogs in each group.The left kidney was preserved and the right kidney was removed in the sham operation group.The model group received autologous heterotopic left kidney transplantation and the right kidney was removed.The experimental operators were all from the same group of physicians.The serum creatinine and urea nitrogen were detected at 8 am on day first,second,third,fifth,seventh,tenth,13th,16th,19th,22th,25th and 28th postoperatively.Two independent samples t-test were used to compare serum creatinine and urea nitrogen of Beagle dogs sham operation group and model group within 28 days after operation.P<0.05 was considered statistically significant.Results One experimental dog in model group was dead due to the serum creatinine continued to rise without any improvement.Euthanasia was performed on the 7th day after kidney transplantation.Anatomy revealed renal artery thrombosis and necrosis of the transplanted kidney.The serum creatinine and urea nitrogen of the other 8 dogs in the model group increased briefly on the second day after surgery,and then gradually decreased and remained stable.The serum creatinine and urea nitrogen of the other 8 dogs in the model group were higher than those in the sham operation group on the first,second,third,fifth,seventh,tenth,13th,16th and 19th days after kidney transplantation,and the differences were statistically significant(t=-4.544,-5.083,-5.171,-4.946,-5.460,-5.345,-3.757,-3.918 and-3.435;t=-4.810,-8.119,-10.379,-12.789,-15.210,-9.983,-5.014,-4.599 and-2.657,all P<0.05).Model group was euthanized after 28 days of observation.Anatomical results showed that the size of the transplanted kidney was about 7 cm×4 cm×2 cm,the texture was moderate,there was intestinal encapsulation around the kidney,there was no thrombosis in the renal artery and internal iliac artery,and the renal vein and external iliac vein returned normally.The blood flow of the reconstructed vessels was good,and no stenosis was observed at the ureter and bladder anastomosis.In the sham operation group,no abnormalities were observed after anatomy,the remaining left kidney showed normal anatomical shape,and the morphological structure and position relationship between renal artery,renal vein and ureter showed normal physiological state.HE staining of transplanted kidney tissue of experimental dogs in model group showed slight changes in renal tissue structure,uniform distribution of glomeruli and no abnormal changes in Baumann′s bursa.The epithelial cells of renal tubules were square or columnar,with abundant cytoplasm,intact lumen structure and no obvious injury.Neutrophils and plasma cells were observed in the renal pelvis of 3 dogs in model group.Conclusions The perioperative management method of autologous kidney transplantation in Beagle dogs was successfully explored through the experiment,which could make the animal model simple,stable and repeatable.