益气活血中药对肾间质纤维化大鼠转化生长因子-β、Smad2和Smad3表达的影响

Effects of Chinese medicine for invigorating qi and promoting blood circulation （益气活血） on the expressions of transforming growth factor-β, Smad2 and Smad3 in rats with renal interstitial fibrosis

目的 探讨益气活血中药对肾间质纤维化大鼠转化生长因子-β（TGF-β）、Smad2和Smad3表达的影响.方法 将36只雄性Wistar大鼠随机分为假手术组、模型组、西药对照组及中药大、中、小剂量组,每组6只.采用单侧输尿管结扎术建立肾间质纤维化动物模型.西药对照组及中药大、中、小剂量组于术前2 d灌胃福辛普利钠（10 mg·kg-1·d-1）及中药提取液（7.2、3.6、1.8 ml·kg-1·d-1）各2 ml;假手术组和模型组给予等量生理盐水.各组于术后3周处死大鼠取肾组织,用实时聚合酶链反应（real-time PCR）和蛋白质免疫印迹法（Western blotting）检测TGF-β、Smad2和Smad3的mRNA和蛋白表达.结果 模型组大鼠肾组织TGF-β、Smad2和Smad3的mRNA和蛋白表达较假手术组均明显升高（P〈0.05或P〈0.01）;益气活血中药各剂量组可以明显抑制模型大鼠肾组织TGF-β、Smad2和Smad3的mRNA和蛋白表达,其中除中药小剂量组对Smad3mRNA的抑制作用较大、中剂量组明显降低外（均P〈0.05）,其余指标各剂量组间比较差异均无统计学意义.结论 益气活血中药可能通过抑制TGF-β/Smad信号通路中TGF-β、Smad2和Smad3等表达起到治疗作用.

Objective To investigate the effects of Chinese medicine for invigorating qi and promoting blood circulation （益气活血）, Yiqi Huoxue formula （YQHXF, 益气活血方）, on the expressions of transforming growth factor-β （TGF-β）, Smad2 and Smad3 in rats with renal interstitial fibrosis. Methods Thirty-six male Wistar rats were randomly divided into six groups： sham operation group, model group, western medicine control group, high-dose, medium dose and low-dose YQHXF groups （each, n = 6）. Unilateral ligation of ureter was carried out to establish the rat model of renal interstitial fibrosis. Two days before the operation, intragastrically, fosinoprii sodium （10 mg ·kg^-1 · d^-1） was given to the western medicine control group, and 2 ml of each of the following doses of YQHXF： 7.2, 3.6, 1.8 ml ·kg^-1 · d^-1 was administered into the high, medium and low dosage groups, respectively. Sham operation group and model group received the same amount of normal saline with the same method. All rats were administered with corresponding drugs for 3 weeks. After the last administration, the animal was sacrificed and its kidney tissue was taken out, the mRNA and protein expressions of TGF-β, Smad 2, Smad3 were measured by the real-time polymerase chain reaction （PCR） and Western blotting. Results The mRNA and protein expressions of TGF-β, Smad2 and Smad3 in model group were higher than those in sham operation group （P〈0. 05 or P〈 0.01）, YQHXF treatment could down-regulate the mRNA and protein expressions of TGF-β, Smad2 and Smad3, and in low-dose YQHXF group, the down-regulation of Smad3 mRNA was more obvious than that in high and medium dosage groups （both P〈0.05）. No significant difference in effect on other indexes was observed among the three YQHXF groups. Conclusion YQHXF might play a role in the treatment of renal interstitial fibrosis via inhibiting the mRNA and protein expressions of TGF-β, Smad2 and Smad3 in the TGF-β/Smad signaling transduction pathway.