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目的:研究益气活血降浊复方对单侧输尿管梗阻(UUO)大鼠肾组织TGF-β1、α-SMA、HGF蛋白及其mRNA表达的影响,探讨本方抑制肾间质纤维化的作用机制。方法:132只雄性SD大鼠以单侧输尿管结扎术建立肾间质纤维化动物模型,造模成功后大鼠随机分为正常组,假手术组,模型组,西药组,益气活血降浊复方大、中、小剂量组。益气活血降浊复方大剂量组给药34.6g/kg,中剂量组17.3g/kg,小剂量组8.6g/kg灌胃给药,模型组和假手术组、正常组灌入相应0.9%氯化钠溶液,西药组灌胃福辛普利钠10g/kg,1次/d。采用免疫组化方法检测肾组织中TGF-β1、α-SMA、HGF蛋白表达,采用Real-time PCR方法检测TGF-β1、α-SMA、HGF mRNA表达。结果:与正常组、假手术组相比,模型组大鼠第1、2、3周肾间质中TGF-β1、α-SMA蛋白表达明显增多(P<0.01),益气活血降浊复方大、中、小剂量组及西药组正常组、假手术组在各个时间点TGF-β1、α-SMA蛋白表达有所增加,但与模型组比较显著降低(P<0.01)。正常组肾小管上皮细胞胞浆中HGF呈弱阳性表达,随着梗阻时间的延长模型组HGF蛋白表达逐渐减少,益气活血降浊复方大、中、小剂量组与模型组相比,HGF蛋白阳性表达显著增加(P<0.01)。与正常组、假手术组相比,造模后第3周模型组肾组织TGF-β1、α-SMA基因高表达(P<0.05,P<0.01),益气活血降浊复方大、小剂量组和福辛普利钠组与模型组比较TGF-β1、α-SMA基因低表达(P<0.05)。造模后第1周,正常组、假手术组HGF基因低表达,益气活血降浊复方大、中、小剂量组与模型组比较HGF基因高表达(P<0.05)。结论:益气活血降浊复方可以促进内源性HGF蛋白和基因的生成,使肾组织α-SMA基因、TGF-β1基因表达降低,抑制肾间质纤维化的发展。
Objective: To study the effect of Yiqi Huoxue Jiangzhuo Compound on the expression of TGF-β1, α-SMA and HGF protein in renal tissue of rats with unilateral ureteral obstruction (UUO), and to explore the mechanism of action of this prescription in inhibiting renal interstitial fibrosis . METHODS: Thirty-two male Sprague-Dawley rats were involved in unilateral ureteral ligation to establish animal model of renal interstitial fibrosis. After successful model establishment, rats were randomly divided into normal group, sham operation group, model group, western medicine group, Yiqi Huoxue Jiangzhuo Compound large, medium and small dose group. Administration of Yiqi Huoxue Jiangzhuo high dose group was given 34.6g / kg, middle dose group 17.3g / kg, low dose group 8.6g / kg by intragastric administration, model group and sham operation group, 0.9% Sodium chloride solution, western medicine group fosinopril sodium 10g / kg, 1 time / d. The expression of TGF-β1, α-SMA and HGF in renal tissue was detected by immunohistochemistry. The mRNA expressions of TGF-β1, α-SMA and HGF were detected by Real-time PCR. Results: Compared with normal group and sham operation group, the expression of TGF-β1 and α-SMA protein in renal interstitium in model group at 1, 2 and 3 weeks were significantly increased (P <0.01) The levels of TGF-β1 and α-SMA in the large, middle and small dose groups and the normal group and the western medicine group were significantly increased at each time point (P <0.01). The expression of HGF in the cytoplasm of normal renal tubular epithelial cells was weakly positive. With the prolongation of the obstruction time, the expression of HGF protein in the model group decreased gradually. Compared with the model group, the HGF protein The positive expression was significantly increased (P <0.01). Compared with normal group and sham operation group, the expression of TGF-β1 and α-SMA gene in model group was higher than that in sham-operated group (P <0.05, P <0.01) Compared with model group, TGF-β1 and α-SMA gene expression was lower (P <0.05). In the first week after model establishment, HGF gene expression was low in normal group and sham operation group, but high in HGF group (P <0.05). Conclusion: Yiqi Huoxue Jiangzhuo Compound can promote the generation of endogenous HGF protein and gene, reduce the expression of α-SMA gene and TGF-β1 gene and inhibit the development of renal interstitial fibrosis.