肾切除联合阿霉素诱导建立家兔局灶节段性肾小球硬化模型

背景:单侧肾切除联合阿霉素诱导的大鼠肾小球硬化模型常用于抗肾小球硬化药物的筛选和评价,但目前很少有此方法诱导兔肾小球硬化模型的报道.目的:建立家兔肾小球硬化动物模型,并观察造模过程中动物的肾脏功能和组织病理学变化.设计、时间及地点:随机对照动物实验,于2008-12/2009-05在解放军成都军区昆明总医院实验动物中心完成.材料:日本大耳家兔25只,雌雄各半,体质量1.75～2.25 kg,用于建立家兔肾小球硬化动物模型,并随机分为正常组10只和模型组15只.方法:模型组30 g/L戊巴比妥钠生理盐水溶液(1 mL/kg)麻醉,开腹摘除左肾.正常组行假手术,不摘除肾脏,余与上同.手术1周后,模型组注射阿霉素5 mg/kg,2 周后注射阿霉素3 mg/kg.正常组同步注射生理盐水.主要观察指标:于实验前、造模后4,6,8周对两组兔进行肾功能相关血液生化指标检测,并留尿液测24 h尿蛋白量.于第2次注药后8周分别两组随机各取1只兔肾组织,进行病理观察.结果:①模型组肾脏略白,质地略韧,表面尚光滑.②模型组光镜下可见细胞外基质增生明显,系膜区扩张,肾球囊壁与肾小球毛细血管拌粘连,小管上皮变性甚至萎缩出现肾间质纤维化,间质炎细胞浸润,呈现弥漫性节段性肾小球硬化等病理特征.③与正常组家兔比较,模型组家兔总蛋白及白蛋白明显下降,三酰甘油、总胆固醇、低密度脂蛋白胆固醇、尿素氮、肌酐均显著升高,模型组家兔尿蛋白含量于实验第4周开始明显升高,在实验期间明显高于对照组,但在第8周逐渐趋向平台.④SPECT示:正常组肾小球总滤过率为92.6 mL/min,模型组为33 mL/min;可见模型组右侧肾脏肾小球滤过率明显下降.结论:单侧肾切除联合阿霉素导致家兔的肾小球硬化形成,并且肾功能及24 h尿蛋白定量有较明显的阶段性变化.

Establishment of focal segmental glomerulosclerosis models induced by nephrectomy combined with adriamycin in rabbits

BACKGROUND: Rat models induced by unilateral nephrectomy combined with adriamycin are widely used in screening anti-drugs of glomerular sclerosis. However, few reports concerning this method on inducing rabbit model of glomerular sclerosisOBJECTIVE: To establish a rabbit model of glomerular sclerosis, and to observe the renal function and histopathological changes during model preparation. DESIGN, TIME AND SETTING: Randomized controlled animal experiment was performed in Laboratory Animal Center of Kunming General Hospital of Chengdu Military Region between December 2008 and May 2009. MATERIALS: Twenty-five Japanese big-ear rabbits, weighing 1.75-2.25 kg, half males and half females, were selected to establish glomerular sclerosis models, and randomly divided into normal (n=10) and model (n=15) groups. METHODS: The left kidney of rabbits in the model group was removed under anesthetized with 30g/L saline solution of sodium pentobarbital (1 mL/kg). Rabbits in the normal group underwent a similar surgical procedure without kidney removing. At 1 week after operation, 5 mg/kg adriamycin was injected into rabbits in the model group, 3 mg/kg adriamycin was reinjected 2 weeks later. Same volume of physiological saline was injected in the normal group. MAIN OUTCOME MEASURES: The renal blood biochemical indexes were detected prior to and at weeks 4, 6, and 8 after model preparation. One nephridial tissue was harvested from each group to undergo pathological observation at week 8 after the second medication. RESULTS: ①Kidney in the normal group presented slightly white, slightly tough texture with smooth surface. ②Pathological features of focal segmental glomerulosclerosis was showed in the model group under a light microscope, presented as extracellular matrix hyperplasia, mesangial region expansion, renal glomerular capillary wall ball mix adhesion, tubule degeneration, or even interstitial fibrosis, as well as interstitial infiltration of inflammatory cells. ③Compared to the normal group, the total protein and albumin was and decreased, triglyceride, total cholesterol, low density lipoprotein cholesterol, urea nitrogen, and creatinine were significantly increased in the model group. The urine protein content of rabbits in the model group was obviously increased at 4 weeks, gradually reached a platform after 8 weeks, which still greater than that of the normal group. ④SPECT showed that the glomerular filtration rate of the model group was notably decreased (33 mL/min) than the normal group (92.6 mL/min).CONCLUSION: The unilateral nephrectomy combined with adriamycin in rabbits results in the formation of glomerular sclerosis, renal function and 24 h urinary protein is more visible in the phase change.