肾病Ⅰ号方醋酸乙酯提取物对单侧输尿管梗阻大鼠肾间质纤维化的影响

Effect of ethyl acetate extract of Shenbing No. 1 Decoction on renal interstitial fibrosis caused by unilateral ureteral obstruction in rats

目的探讨肾病I号方醋酸乙酯提取物(ethyl acetate extract of Shenbingyihao Decoction,EASD)对肾纤维化大鼠转化生长因子-β1(TGF-β1)、α-平滑肌肌动蛋白(α-SMA)表达的影响。方法 50只SD大鼠随机分为假手术组、模型组、洛丁新(洛丁新片1.67 mg/kg)组、肾病I号方(生药18.75 g/kg)组、EASD(EASD,300 mg/kg)组,除假手术组外,其他各组采用单侧输尿管结扎的方法制备单侧输尿管梗阻(UUO)大鼠模型。各组ig给药14 d后处死大鼠,采集血清,检测尿素氮(BUN)和血肌酐(Scr)水平,取肾组织行HE和Masson染色评价肾小管间质纤维化损伤程度,采用免疫组织化学染色方法和实时荧光定量PCR(RT-PCR)法检测TGF-β1、a-SMA蛋白及m RNA的表达情况。结果与假手术组比较,模型组和3个给药组大鼠血清Scr、BUN升高(P<0.05),间质纤维化程度加重(P<0.05),肾组织TGF-β1、a-SMA表达明显增多(P<0.05)。与模型组比较,3个给药组大鼠的血清Scr、BUN下降(P<0.05),肾组织TGF-β1、a-SMA表达下降(P<0.05)。结论肾病I号方及EASD能够延缓肾纤维化的发生发展,其作用机制可能是通过下调TGF-β1、a-SMA的表达,从而达到保护肾脏的作用。

Objective To investigate the effect of ethyl acetate extract of Shenbing No. 1 Decoction（EASD） on the expression of TGF-β1 and a-SMA in rats with renal fibrosis. Methods Totally 50 SD rats were randomly divided into Sham operation group, unilateral ureteral obstruction（UUO） model group, Lotensin treated group（1.67 mg/kg）, Shenbing No. 1 Decoction group（18.75 g/kg） and EASD group（300 mg/kg）. A rat model of renal fibrosis was induced by unilateral ureteral ligation. The rats were executed after 14 d. The contents of 24 h urinary protein（24h UP）, serum blood urea nitrogen（BUN）, and serum creatitine（Scr） were tested. Renal tubular interstitial fibrosis was evaluated by HE staining and Masson staining. The protein expression levels of TGF-β1 and a-SMA were observed by immunohistochemical method. Quantitative real-time PCR was adopted to inspect the m RNA expression of a-SMA and TGF-β1. Results Compared with Sham-operation group, in rats of other groups, the serum levels of BUN and SCr increased（P〈0.05）, renal interstitial fibrosis degree increased, and protein expression levels of TGF-β1 and a-SMA increased in the renal tissue（P〈0.05）. Compared with UUO model group, in rats of other groups, the serum levels of BUN and SCr decreased（P〈0.05） and protein expression levels of TGF-β1 and a-SMA decreased（P〈0.05）. There was no significant difference among three treatment groups（P〉0.05）. Conclusion Shenbing No. 1 Decoction and EASD may ameliorate renal interstitial fibrosis by down-regulating the expression levels of TGF-β1 and a-SMA.