商陆及其不同炮制品对阿霉素肾病大鼠的作用机制分析

Analysis of Mechanism of Phytolaccae Radix and Its Processed Products on Doxorubicin-induced Nephropathy in Rats

目的:比较商陆及其不同炮制品对阿霉素肾病大鼠的保护作用,并初步探究其作用机制。方法:采用一次性尾静脉注射盐酸阿霉素的方法建立大鼠肾病模型。通过HPLC-ELSD测定商陆及其不同炮制品中商陆皂苷甲(Es A)的含量。利用酶联免疫吸附试验(ELISA)测定各组大鼠血清总蛋白(TP),白蛋白(Alb),总胆固醇(TC),血清肌酐(SCr),尿素氮(BUN)和尿蛋白(UP)的含量。通过实时荧光定量聚合酶链式反应(Real-time PCR)和免疫组织化学法测定大鼠肾脏组织转化生长因子-β(TGF-β) mRNA和蛋白的表达。结果:与空白组比较,模型组大鼠血清TP和Alb含量显著降低(P <0. 05);TC,SCr和BUN含量显著升高(P <0. 05),尿中UP含量显著升高(P <0. 05)。商陆生品、炮制品和Es A均能不同程度的升高模型大鼠血清TP和Alb的表达,降低TC,SCr和BUN以及尿中UP的含量;其中醋炙品的作用最为显著。Real-time PCR和免疫组化结果均显示,模型组大鼠肾脏组织TGF-β表达较空白组显著升高(P <0. 01),而醋炙商陆、醋煮商陆和清蒸商陆均能显著降低模型大鼠肾脏组织TGF-β的表达(P <0. 05,P <0. 01)。结论:商陆及其炮制品均能不同程度的改善阿霉素肾病模型大鼠的症状,其中醋炙品的作用最强,并且该作用可能与降低肾脏组织的TGF-β表达有关。

Objective:To compare the protective effect of Phytolaccae Radix and its processed products on nephropathy induced by doxorubicin(DOX)in rats,and explore its mechanism.Method:A rat model of nephropathy was established by a single tail intravenous injection of DOX hydrochloride.Content of esculentoside A(EsA)in Phytolaccae Radix and its processed products was determined by HPLC-ELSD.Contents of serum total protein(TP),albumin(Alb),urea nitrogen(BUN),serum creatinine(SCr),total cholesterol(TC)and urine protein(UP)were determined by enzyme-linked immunosorbent assay(ELISA).The mRNA and protein expression of transforming growth factor-β(TGF-β)in renal tissue of rats was examined by real-time quantitative polymerase chain reaction(Real-time PCR)and immunohistochemistry.Result:A single intravenous injection of DOX could induce a severe nephrotic syndrome associated with decreased serum TP,Alb and elevated serum BUN,SCr,TC,and a high urinary excretion of protein(P<0.05).Expression of BUN,SCr,TC in serum and UP in urine of model rats could be decreased by Phytolaccae Radix and its processed products to some degree,expression of TP and Alb in serum of model rats could be increased by Phytolaccae Radix and its processed products to some degree,vinegar processed products had the most significant effect.Expression of TGF-βin renal tissue of model group rats was significantly higher than that of blank group(P<0.01),which could be significantly reversed by Phytolaccae Radix processed with vinegar,Phytolaccae Radix boiled with vinegar,Phytolaccae Radix steamed with water(P<0.05,P<0.01).Conclusion:Phytolaccae Radix and its processed products can improve the symptoms of DOX nephropathy model rats in different degrees,among which the vinegar prepared products have the strongest effect,and this effect may be related to the reduction of TGF-βexpression in renal tissue.