左肾切除术联合链脲佐菌素腹腔注射诱导糖尿病肾病大鼠模型的建立

Establishment of rat model of diabetic nephropathy induced by left nephrectomy combined with intraperitoneal injection of streptozotocin

目的探讨左肾切除术联合链脲佐菌素腹腔注射诱导糖尿病肾病(DN)大鼠模型的建立。方法健康雄性SD大鼠共16只,随机分为3组,对照组(5只)、假手术组(5只)和模型组(6只)。适应性喂养3 d后,模型组行左侧肾脏切除术,术后1周单次腹腔注射60 mg/kg链脲佐菌素,建立DN大鼠模型;假手术组做手术切口,但不切除左肾,术后给予等体积的柠檬酸缓冲液腹腔注射;对照组小鼠不做任何处理。以造模后72 h内2次随机血糖大于16.7 mmol/L为糖尿病模型成立标准。在成模后3个月时所有大鼠留取血液、尿液及肾脏标本,检测血尿生化指标,HE、PAS、Masson染色及电镜观察肾脏病理特点。结果与对照组及假手术组比较,模型组大鼠体质量增加缓慢,后期体质量呈负增长趋势,饮水量明显增加,且出现糖尿病视网膜病变,24h尿量、24h尿蛋白定量及尿微量白蛋白定量显著增加,血肌酐、尿素氮水平升高,HE、PAS、Masson染色提示:模型组大鼠肾小球基底膜增厚,肾小管空泡样变性,血管内膜增厚,部分大鼠出现肾间质纤维化。电镜提示:肾小球基底膜增厚,部分足细胞足突融合或缺失。结论左肾切除术联合链脲佐菌素腹腔注射能够建立可靠的DN大鼠模型。

Objective To investigate the establishment of a rat model of diabetic nephropathy induced by intraperitoneal injection of streptozotocin and left nephrectomy.Methods Sixteen healthy male SD rats were randomly divided into 3 groups：normal control group（N group,5 rats）,sham operation group（SO group,5 rats）,and model group（M group,6 rats）.After 3 days of sexual feeding,M group underwent left kidney resection,and a single intraperitoneal injection of 60 mg/kg streptozotocin was performed one week after surgery to establish a diabetic nephropathy rat model.Two times at 72 h after modeling,randomized blood glucose levels greater than 16.7 mmol/L were criteria for the establishment of a diabetic model.Blood,urine,and kidney specimens were collected at the 3 rd month after modeling,and blood and urine biochemical parameters were measured.HE,PAS,Masson staining,and electron microscopy were used to observe the pathological features of the kidneys.Results As compared with N group and SO group,the weight gain in M group was slowed down,the body weight showed a negative growth trend at the later stage,the drinking water amount increased significantly,and diabetic retinopathy occurred.The urine volume,urinary protein and urinary albumin quantification were significantly increased at 24 h in M group.Blood creatinine and blood urea nitrogen levels were elevated in M group.HE,PAS,and Masson staining indicated that in M group,glomerular basement membrane and vascular intima were thickened,tubular vacuolar degeneration occurred,and some rats developed kidney Interstitial fibrosis.Electron microscopy suggested that the glomerular basement membrane was thickened and some podocyte foot processes were fused or absent.Conclusions A left kidney nephrectomy combined with intraperitoneal injection of streptozotocin can establish a reliable rat model of diabetic nephropathy.