嘌呤霉素氨基核苷诱发肾病综合征大鼠模型的改良制作研究

Studies of Improved Rat Model with Nephrotic Syndrome Induced by Puromycin Aminonucleoside

目的:在嘌呤霉素氨基核苷模型经典制作方法的基础上进行多次尾静脉追加,制作更适合慢性肾脏病药理学研究的实验性大鼠模型。方法:Wistar大鼠42只,6周龄,雄性,随机分为对照组、颈静脉给药组和追加给药组,每组14只,各组分别于造模后第30天和第56天处死动物。颈静脉给药组行颈静脉插管术缓慢推注嘌呤霉素氨基核苷(40 mg/kg),追加给药组在颈静脉给药(40 mg/kg)后第13、16、19天再进行3次小剂量尾静脉追加(5 mg/kg)。对照组给等容积0.9%生理盐水。造模后第5、10、15、20、28、42、56天时检测24 h尿蛋白。实验结束时对比两组模型的肾脏组织病理改变和血清总蛋白(TP)、白蛋白(Alb)、三酰甘油(TG)、胆固醇(TC)、肌酐(Scr)和尿素氮(BUN)水平。结果:颈静脉给药组在造模后第5天出现蛋白尿,10 d左右尿蛋白达到高峰(均值264.1 mg/24 h),随后尿蛋白急剧下降,约在第20天降至接近正常水平。追加给药组尿蛋白出现一个持续增高的平台期(均值维持在略高于150 mg/24 h的水平),末次追加后约10 d(颈静脉给药后第28天)开始缓慢下降,于第56天时接近正常。光镜所见:在30 d和56 d时,追加给药组大鼠的肾小球局灶节段性硬化和间质纤维化、炎性浸润等组织病理学指标较颈静脉给药组改变显著,其中两组之间间质纤维化的差异有统计学意义(P<0.05)。第30天血清生化结果显示,追加给药组大鼠TG、TC(P<0.05)、Scr(P<0.05)升高较明显,而颈静脉给药组变化则不明显。结论:对嘌呤霉素氨基核苷经典模型进行多次小剂量尾静脉追加,可以造成大鼠慢性肾脏病变。改良后的模型能够节约研究时限和研究成本,更适用于开展慢性肾脏病药理学研究。

Objective：In order to explore a more stable and reliable rat model with chronic kidney disease by vena caudalis superadditions of puromycin aminonucleoside（PAN） in rats after classic PAN procedure.Methods：Forty-two male Wistar rats,6-weeks old,were randomely divided into the control group,the jugular vein injection group and the superaddition group.Each group comprised of 14 rats which were sacrificed at the 30th and the 56th days after model established.Rats in the jugular vein injection group were performed with single jugular vein injection of PAN at a dose of 40 mg/kg;while rats in the superaddition group were primarily performed with jugular vein injection of PAN at the same dose followed with vena caudalis injection of PAN at a dose of 5 mg/kg for 3 times,respectively at 13th,16th and 19th day.Rats in the control group were injected with 0.9% normal saline at the same volume.The changes of urine protein were examined at 5th、10th、15th、20th、28th、42nd and 56th day.At the end of the experiment,the serum triglycerin,cholesterol,creatinine and urea nitrogen levels between the two groups were examined,and the pathological alteration of renal tissue were compared between the two groups.Results：In the jugular vein injection group,albuminuria in all rats was observed at the 5th day that reached its peak level around the 10th day and decreased rapidly into normal range around the 20th day.While the elevated albuminuria level in the superaddition group after three low dose superadditions retained in plateau period（mean over 150 mg/24 h）.Ten days after last superaddition（the 19th day after model established）,the albuminuria level in this group decreased gradually into normal range around the 56th day.At the 30th and the 56th days,the changes of focal segmental glomerulosclerosis,interstitial fibrosis and inflammatory infiltration in the renal tissues were more prominent in the superaddition group than those in the jugular vein injection group,and the intersitial fibrosis changes of the two group were obvious significance（P〈0.05）.At the 30th day,the biochemistry parameters,such as triglycerin,total cholesterol（P〈0.05） and serum creatinine level（P〈0.05）,were increased in the superaddition group while those alterations in the jugular vein injeciton group were not obvious.Conclusion：The method of vena caudalis superadditions of PAN at low dosage is successful in inducing chronic kidney disease on the basis of the classic PAN rat model.The improved model is more suitable for pharmacologic research on chronic kidney disease due to more economical in time and cost.