柴苓归芪汤加蚓激酶对实验性肾病综合征大鼠肾功能的保护作用

1、柴苓归芪汤加蚓激酶对实验性肾病综合征大鼠肾功能的保护作用              作者：张芬芳，王喜栋，范焕芳，陈志强，尹智炜，张伟【关键词】  柴苓归芪汤    摘要  目的：探讨柴苓归芪汤加蚓激酶对实验性肾病综合征大鼠肾功能的保护作用。方法：采用尾静脉一次性注射阿霉素5 mg/kg制作肾病大鼠模型，随机分为模型组、辛伐他汀组、柴苓归芪组，并设正常对照组。造模1周后，予以药物干预，共用药5周。采用三氯乙酸法测定大

2、鼠24 h尿蛋白含量；全自动生化分析仪检测血清总蛋白（total protein, TP）、白蛋白（albumin, Alb）、总胆固醇（total cholesterol, TC）、甘油三酯（triglycerides, TG）、高密度脂蛋白（highdensity lipoprotein, HDL）、低密度脂蛋白（lowdensity lipoprotein, LDL）、肌酐（creatinine, Cr）及血尿素氮（blood urea nitrogen, BUN）。光镜、电镜下观察肾组织病理形态学变化。结果：模型组与正常对照组比较，24 h尿蛋白定量及血清TC、TG、LDL、Cr、BU

3、N均明显升高（P0.05或P0.01)；血清TP、Alb、HDL则明显降低（P0.01)。柴苓归芪汤组24 h尿蛋白定量及血清TC、TG、LDL、Cr、BUN均明显低于模型组（P0.05或P0.01)；而血清TP、Alb和HDL则明显高于模型组（P0.05或P0.01)。肾组织病理形态学观察：模型组光镜下部分肾小球呈局灶节段性硬化，近曲小管上皮细胞变性肿胀，部分小管腔内可见蛋白管型，间质可见散在的纤维增生；电镜下肾小球上皮细胞足突广泛融合。柴苓归芪汤组未出现肾小球硬化，肾小管间质病变明显减轻，肾小球上皮细胞足突少量融合。结论：柴苓归芪汤能够降低大鼠尿蛋白，调节脂质代谢紊乱，保护肾脏功能，延缓肾

4、脏疾病进展。关键词  柴苓归芪汤; 阿霉素肾病; 蛋白尿; 脂质代谢; 高脂血症; 肾组织; 病理形态学Protective effects of Chailing Guiqi Decoction combined with lumbrukinase on renal function in rats with adriamycin nephropathyABSTRACT  Objective: To study the protective efffects of Chailing Guiqi Decoction (CLGQD) combined with lumbruk

5、inase on renal function in rats with adriamycin nephropathy. Methods: Thirtysix SD rats were randomly divided into four groups: normal control group, untreated group, simvastatintreated group and CLGQD treated group. Adriamycin nephropathy was induced by intravenous injection with 5 mg/kg adriamycin

6、. After sevenday treatment, quantitative measurement of 24h urine protein was determined with trichloroacetic acid, and serum total protein (TP), albumin (Alb), total cholesterol (TC), triglycerides (TG), highdensity lipoprotein (HDL), lowdensity lipoprotein (LDL), creatinine (Cr) and blood urea nit

7、rogen (BUN) were assessed using automatic biochemistry analyzer. The pathomorphological changes of renal tissues were observed with light and electron microscopes. Results: In the untreated group, the 24h urine protein excretion, serum TC, TG, LDL, Cr and BUN were significantly higher than those in

8、the normal control group (P0.05 or P0.01), while the serum TP, Alb, HDL were significantly lower than those in the normal control group (P0.01). In the CLGQDtreated group, the 24h urine protein excretion, serum TC, TG, LDL, Cr and BUN were significantly lower as compared with those in the untreated

9、group (P0.05 or P0.01), while the serum TP, Alb and HDL were significantly higher as compared with those in the untreated group (P0.05 or P0.01). The pathomorphological findings of the renal tissues under the light microscope in the untreated group showed focal segmental glomerulosclerosis in a few

10、of glomerulus, degenerated and swelled proximal tubular epithelial cells, proteins in cast formation in some renal tubules and scattered fibrosis in interstitial tissues of the kidney, while the electron microscope images showed the fusion of foot processes in glomerular epithelial cells. The pathom

11、orphological changes in the CLGQDtreated group were slighter than those in the untreated group. Conclusion: CLGQD combined with lumbrukinase can reduce proteinuria, regulate lipid metabolism, protect renal function, and delay progressive renal damage in rats.KEY WORDS  Chailing Guiqi Decoction;

12、 adriamycin nephropathy; proteinuria; lipid metabolism; hyperlipidemia; renal tissue; pathomorphology阿霉素肾病（adriamycin nephropathy, AN）大鼠模型是经典的实验性肾病综合征模型之一，初期的病理改变类似于人的微小病变型肾病，进一步发展可形成局灶节段性肾小球硬化1。本实验采用此模型，观察柴苓归芪汤对大鼠血、尿生化指标及肾组织病理形态学的影响，并以他汀类降脂药物辛伐他汀作为对照，重点观察该方对尿蛋白及脂质代谢的影响，探讨该方保护肾功能及防止肾功能衰竭发生的作用机制。1

13、60; 材料与方法1.1  材料1.2  分组、造模及药物处理  所有实验动物予以普通饲料适应性喂养1周后，置于清洁代谢笼内取尿以检测尿蛋白和尿红细胞，全部为阴性。36只大鼠随机分为正常对照组（6只）、模型组（10只）、辛伐他汀组（10只）和柴苓归芪组（10只）。模型组、辛伐他汀组和柴苓归芪组予以尾静脉注射阿霉素（5 mg/kg），正常对照组注射等量生理盐水。造模1周后开始灌胃，辛伐他汀组给予辛伐他汀1 mgkg1d1，柴苓归芪组给予柴苓归芪汤煎剂27 gkg1d1，用药剂量均为成人的15倍；蚓激酶按25 mg/kg，2次/d给药3。正常对照组和模型组均灌服等量蒸

14、馏水。共用药5周。1.3  检测指标与方法1.4  肾组织病理形态学观察  实验第7周末处死动物，取部分新鲜肾组织分别行光镜和电镜观察。光镜观察：肾组织用4%多聚甲醛溶液予以固定，酒精梯度脱水，二甲苯中透明，石蜡包埋，制成厚度2 m的切片，行高碘酸希夫氏染色（periodic acidSchiff stain, PAS）和Masson染色，观察肾小球及肾小管间质的变化。电镜观察：肾组织用4%戊二醛固定，1%锇酸磷酸钠缓冲液后固定，丙酮梯度脱水，815环氧树脂包埋，Leica UCT超薄切片机切片，醋酸双氧铀和柠檬酸铅双重染色，在日立H7500透射电镜下观察。1.5

15、  统计学方法  采用SPSS 10.0软件进行统计学分析，计量资料用x±s表示，采用单因素方差分析。2  结果 2.1  大鼠一般情况  正常对照组大鼠未出现死亡；辛伐他汀组大鼠于实验第6周死亡1只；模型组和柴苓归芪组于实验第7周各死亡1只，可能系感染所致，尾部的感染可能与注射部位阿霉素渗漏引起局部缺血坏死有关。2.2  对24 h尿蛋白定量的影响  实验第2周末各组大鼠均未出现明显的蛋白尿。模型组第3周末出现蛋白尿（24 h尿蛋白定量100 mg），第4周末蛋白尿进一步加重，第5周末达高峰，第6、7周末蛋白尿稍

16、减轻，其第3、4、5、6、7周末24 h尿蛋白定量与正常对照组比较均有统计学意义（P0.01)。辛伐他汀组和柴苓归芪组第3周末的尿蛋白定量与模型组相比虽有下降，但无统计学意义；第4、5周末尿蛋白定量明显下降，与模型组比较有统计学意义（P0.05或P0.01)；第6、7周末模型组的尿蛋白定量虽有下降，但与辛伐他汀组和柴苓归芪组比较，仍有统计学意义（P0.05或P0.01)。实验中，虽然柴苓归芪组24 h尿蛋白定量始终略高于辛伐他汀组，但两组比较无统计学意义。见表1。2.3  对血清TP、Alb、BUN、Cr的影响  实验第7周末模型组的血清TP、Alb值较正常对照组明显降低（P0.05或P0.01）。辛伐他汀组和柴苓归芪组血清TP、Alb值较模型组显著升高（P0.05或P0.01）。柴苓归芪组血清TP、Alb值略高于辛伐他汀组，但两者比较无统计学意义。实验第7周末，模型组的血清BUN、Cr值与正常对照组比较有显著性升高（P0.05或P0.01）。辛伐他汀组、柴苓归芪组血清BUN、Cr值与模型组比较均有