柴苓歸芪湯加蚓激酶對實(shí)驗(yàn)性腎病綜合征大鼠腎功能的保護(hù)作用

1、柴苓歸芪湯加蚓激酶對實(shí)驗(yàn)性腎病綜合征大鼠腎功能的保護(hù)作用              作者：張芬芳，王喜棟，范煥芳，陳志強(qiáng)，尹智煒，張偉【關(guān)鍵詞】  柴苓歸芪湯    摘要  目的：探討柴苓歸芪湯加蚓激酶對實(shí)驗(yàn)性腎病綜合征大鼠腎功能的保護(hù)作用。方法：采用尾靜脈一次性注射阿霉素5 mg/kg制作腎病大鼠模型，隨機(jī)分為模型組、辛伐他汀組、柴苓歸芪組，并設(shè)正常對照組。造模1周后，予以藥物干預(yù)，共用藥5周。采用三氯乙酸法測定大

2、鼠24 h尿蛋白含量；全自動(dòng)生化分析儀檢測血清總蛋白（total protein, TP）、白蛋白（albumin, Alb）、總膽固醇（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）。光鏡、電鏡下觀察腎組織病理形態(tài)學(xué)變化。結(jié)果：模型組與正常對照組比較，24 h尿蛋白定量及血清TC、TG、LDL、Cr、BU

3、N均明顯升高（P0.05或P0.01)；血清TP、Alb、HDL則明顯降低（P0.01)。柴苓歸芪湯組24 h尿蛋白定量及血清TC、TG、LDL、Cr、BUN均明顯低于模型組（P0.05或P0.01)；而血清TP、Alb和HDL則明顯高于模型組（P0.05或P0.01)。腎組織病理形態(tài)學(xué)觀察：模型組光鏡下部分腎小球呈局灶節(jié)段性硬化，近曲小管上皮細(xì)胞變性腫脹，部分小管腔內(nèi)可見蛋白管型，間質(zhì)可見散在的纖維增生；電鏡下腎小球上皮細(xì)胞足突廣泛融合。柴苓歸芪湯組未出現(xiàn)腎小球硬化，腎小管間質(zhì)病變明顯減輕，腎小球上皮細(xì)胞足突少量融合。結(jié)論：柴苓歸芪湯能夠降低大鼠尿蛋白，調(diào)節(jié)脂質(zhì)代謝紊亂，保護(hù)腎臟功能，延緩腎

4、臟疾病進(jìn)展。關(guān)鍵詞  柴苓歸芪湯; 阿霉素腎病; 蛋白尿; 脂質(zhì)代謝; 高脂血癥; 腎組織; 病理形態(tài)學(xué)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阿霉素腎?。╝driamycin nephropathy, AN）大鼠模型是經(jīng)典的實(shí)驗(yàn)性腎病綜合征模型之一，初期的病理改變類似于人的微小病變型腎病，進(jìn)一步發(fā)展可形成局灶節(jié)段性腎小球硬化1。本實(shí)驗(yàn)采用此模型，觀察柴苓歸芪湯對大鼠血、尿生化指標(biāo)及腎組織病理形態(tài)學(xué)的影響，并以他汀類降脂藥物辛伐他汀作為對照，重點(diǎn)觀察該方對尿蛋白及脂質(zhì)代謝的影響，探討該方保護(hù)腎功能及防止腎功能衰竭發(fā)生的作用機(jī)制。1

13、60; 材料與方法1.1  材料1.2  分組、造模及藥物處理  所有實(shí)驗(yàn)動(dòng)物予以普通飼料適應(yīng)性喂養(yǎng)1周后，置于清潔代謝籠內(nèi)取尿以檢測尿蛋白和尿紅細(xì)胞，全部為陰性。36只大鼠隨機(jī)分為正常對照組（6只）、模型組（10只）、辛伐他汀組（10只）和柴苓歸芪組（10只）。模型組、辛伐他汀組和柴苓歸芪組予以尾靜脈注射阿霉素（5 mg/kg），正常對照組注射等量生理鹽水。造模1周后開始灌胃，辛伐他汀組給予辛伐他汀1 mgkg1d1，柴苓歸芪組給予柴苓歸芪湯煎劑27 gkg1d1，用藥劑量均為成人的15倍；蚓激酶按25 mg/kg，2次/d給藥3。正常對照組和模型組均灌服等量蒸

14、餾水。共用藥5周。1.3  檢測指標(biāo)與方法1.4  腎組織病理形態(tài)學(xué)觀察  實(shí)驗(yàn)第7周末處死動(dòng)物，取部分新鮮腎組織分別行光鏡和電鏡觀察。光鏡觀察：腎組織用4%多聚甲醛溶液予以固定，酒精梯度脫水，二甲苯中透明，石蠟包埋，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，觀察腎小球及腎小管間質(zhì)的變化。電鏡觀察：腎組織用4%戊二醛固定，1%鋨酸磷酸鈉緩沖液后固定，丙酮梯度脫水，815環(huán)氧樹脂包埋，Leica UCT超薄切片機(jī)切片，醋酸雙氧鈾和檸檬酸鉛雙重染色，在日立H7500透射電鏡下觀察。1.5

15、  統(tǒng)計(jì)學(xué)方法  采用SPSS 10.0軟件進(jìn)行統(tǒng)計(jì)學(xué)分析，計(jì)量資料用x±s表示，采用單因素方差分析。2  結(jié)果 2.1  大鼠一般情況  正常對照組大鼠未出現(xiàn)死亡；辛伐他汀組大鼠于實(shí)驗(yàn)第6周死亡1只；模型組和柴苓歸芪組于實(shí)驗(yàn)第7周各死亡1只，可能系感染所致，尾部的感染可能與注射部位阿霉素滲漏引起局部缺血壞死有關(guān)。2.2  對24 h尿蛋白定量的影響  實(shí)驗(yàn)第2周末各組大鼠均未出現(xiàn)明顯的蛋白尿。模型組第3周末出現(xiàn)蛋白尿（24 h尿蛋白定量100 mg），第4周末蛋白尿進(jìn)一步加重，第5周末達(dá)高峰，第6、7周末蛋白尿稍

16、減輕，其第3、4、5、6、7周末24 h尿蛋白定量與正常對照組比較均有統(tǒng)計(jì)學(xué)意義（P0.01)。辛伐他汀組和柴苓歸芪組第3周末的尿蛋白定量與模型組相比雖有下降，但無統(tǒng)計(jì)學(xué)意義；第4、5周末尿蛋白定量明顯下降，與模型組比較有統(tǒng)計(jì)學(xué)意義（P0.05或P0.01)；第6、7周末模型組的尿蛋白定量雖有下降，但與辛伐他汀組和柴苓歸芪組比較，仍有統(tǒng)計(jì)學(xué)意義（P0.05或P0.01)。實(shí)驗(yàn)中，雖然柴苓歸芪組24 h尿蛋白定量始終略高于辛伐他汀組，但兩組比較無統(tǒng)計(jì)學(xué)意義。見表1。2.3  對血清TP、Alb、BUN、Cr的影響  實(shí)驗(yàn)第7周末模型組的血清TP、Alb值較正常對照組明顯降低（P0.05或P0.01）。辛伐他汀組和柴苓歸芪組血清TP、Alb值較模型組顯著升高（P0.05或P0.01）。柴苓歸芪組血清TP、Alb值略高于辛伐他汀組，但兩者比較無統(tǒng)計(jì)學(xué)意義。實(shí)驗(yàn)第7周末，模型組的血清BUN、Cr值與正常對照組比較有顯著性升高（P0.05或P0.01）。辛伐他汀組、柴苓歸芪組血清BUN、Cr值與模型組比較均有