阿霉素诱发局灶节段性肾小球硬化肾病大鼠模型的建立

目的 建立临床表现和病理特点与人类肾小球疾病相似的阿霉素肾病大鼠模型.方法 将40只SD大鼠完全随机分为造模组(32只)和对照组(8只),造模组完全随机分为首次注射阿霉素后2周组、4周组、8周组、12周组,每组各8只.造模组大鼠尾静脉分2次注射阿霉素,第1次注射按6 mg/kg给药,第2次子1周(7 d)后按4 mg/kg给药；对照组按照造模组方法经大鼠尾静脉2次注射等量生理盐水.检测各组大鼠24h尿蛋白量、血清总蛋白、白蛋白、CH、TG、BUN、Cr的变；双抗体夹心酶联免疫吸附方法检测各组大鼠血清基质金属蛋白酶( MMP)2、MMP-9、转化生长因子(TGF) -β1；免疫组化方法检测各组大鼠纤维连接蛋白(FN)和层粘连蛋白(LN)的表达化；光镜、电镜观察2组大鼠肾脏病理变化.结果 造模组24h尿蛋白定量明显增加,8周末达高峰.造模组大鼠2周、4周、8周和12周组与对照组相比,血清总蛋白、白蛋白明显降低[(50.7±3.6)、(42.2±3.4)、(40.4±3.1)、(41.7±3.3)g/L比(62.8±2.8) g/L、(22.2±1.8)、(16.4±1.7)、(14.8±2.1)、(13.0±2.2)g/L比(29.0±1.3)g/L]；TG、CH明显升高[(5.2±1.4)、(6.1±1.5)、(7.3±1.5)、(7.2±1.2)mmol/L比(0.6±0.3)mmol/L,(3.8±1.7)、(5.6±1.9)、(8.6±2.7)、(9.6±1.6) mmol/L比(1.3±0.1)mmol/L];MMP-2、MMP-9降低[(151.4±2.4)、(100.5±3.1)、(76.5±3.6)、(83.7±3.1)μg/L;(73.4±2.7)、(58.9±2.2)、(35.4±2.0)、(33.3±1.5) μg/L],血清TGF-β1含量升高；肾组织FN、LN的表达和分布范围均明显增加.造模12周在可见部分肾小球出现局灶节段性硬化.结论 采用阿霉素首次大剂量(6mg/kg)冲击诱导,1周后小剂量(4mg/kg)给药的方法,大鼠在12周末可见部分肾小球出现局灶节段性硬化.

The research of adriamycin induced focal segmental glomerulosclerosis in rat

Objective To establish the model of rat adriamycin induced nephritis (AIN)similar to the human renal glomerular disease in patho and amynology.Methods Forty Sprague Dawley rats were randomly divided into model(32) and control group (8).They were randomly divided into the first injection of adriamycin after 2 weeks,4 weeks,8 weeks and 12 weeks.Rat tail of model group vein were injected of adriamycin for two times;control group was given two injections of the same amount of normal saline through the tail veins of rats according to the method with the model group.The level of 24 hour urinary protein,serum total protein(TP),albumin(ALB),cholesterin (CH),triglyeride ( TG),serum urea nitrogen (BUN) and serum creatinine ( Cr ) were measured.The renal pathological changes were observed by light microscope and electron microscope.The expression of fibronectin(FN)and laminin (LN)was analyzed by immunohistochemistry.Matrix metalloproteinases(MMP)-2,MMP-9 and transforming grouth factor(TGF)-β1 were detected by enzyme linked immunosorbent assay.Results Compared with control group,the level of 24 hour urinary protein in AIN model at 2th,4th,8th and 12th week increased( P ＜ 0.01 ),TP and ALB were obviously decreased[ (50.7 ± 3.6),(42.2 ± 3.4),(40.4 ± 3.1 ),(41.7 ± 3.3 ) g/L vs (62.8 ±2.8)g/L;(22.2 ±1.8),(16.4 ±1.7),(14.8 ±2.1),(13.0 ±2.2)g/L vs (29.0 ± 1.3)g/L]; TG and CH in AIN model group were obviously increased [ (5.2 ± 1.4),(6.1 ± 1.5 ),(7.3 ± 1.5 ),(7.2 ± 1.2 ) mmol/L vs (0.6 ±0.3)mmol/L;(3.8 ±1.7),(5.6 ±1.9),(8.6 ±2.7),(9.6 ±1.6)mmol/L vs (1.3 ±0.1)mmol/L] ;focal segmental glomerulosclerosis( FSGS)were observed in 12th week group; The level of TGF-β1 in serum were obviously increased,the level of MMP-2 and M MP-9 reduced [ ( 151.4 ± 2.4 ),( 100.5 ± 3.1 ),(76.5 ± 3.6),( 83.7 ±3.1 ) μg/L; (73.4 ± 2.7),(58.9 ± 2.2 ),( 35.4 ± 2.0),( 33.3 ± 1.5 ) μg/L ] ; the expression and distribution of FN,LN increased significantly.Conclusion After the first large doses of adriamycin (6 mg/kg)shockb is induced,plus one week after low dose (4 mg/kg) administration in rats,the rats appear visiblely part of giomerular appearance of FSGS at the end of 12 weeks.