二黄益肾汤调控TGF-β1/Smad信号通路治疗慢性肾功能衰竭研究

Mechanism of Erhuang Yishen Decoction in treating chronic renal failure via inactivation of the TGF-β1/Smad signal pathway

关键词:  肾功能衰竭  血肌酐  转化生长因子-β1  肾间质

Key words:  renal failure  serum creatinine  transforming growth factor-β1  renal interstitium

摘要:

[目的] 观察二黄益肾汤对5/6肾切除大鼠肾组织血清尿素氮（BUN）、血清肌酐（Scr）、转化生长因子-β1（TGF-β1）、Smad3、Smad4、Smad7 mRNA及蛋白表达的影响。[方法] 健康SPF级雄性SD大鼠，采用5/6肾切除方式建立慢性肾功能衰竭模型。造模成功后，随机分为模型组、尿毒清组和二黄益肾汤组，各给药组分别给予相应药物进行灌胃治疗，时间为12周。实验结束后，检测BUN、Scr含量，苏木精-伊红（HE）染色分析病理改变。采用实时定量-聚合酶链反应（RT-PCR）和蛋白免疫印迹（Western Blot）方法检测TGF-β1、Smad3、Smad4和Smad7 mRNA及蛋白表达量。[结果] 5/6肾切除方法可以成功建立慢性肾功能衰竭动物模型。与模型组比较，二黄益肾汤组与尿毒清组能够降低Scr、BUN含量，差异具有统计学意义（P<0.01）。二黄益肾汤组和尿毒清组肾间质内有少量炎性细胞浸润及纤维组织增生，纤维化面积缩小，能够改善肾功能衰竭肾纤维化病变程度。基因和蛋白表达量分析结果显示，与模型组比较，二黄益肾汤组和尿毒清组TGF-β1、Smad3、Smad4表达呈降低趋势，差异有统计学意义（P<0.05），Smad7的表达呈升高趋势，差异有统计学意义（P<0.05）。[结论] 二黄益肾汤能够改善慢性肾功能衰竭大鼠肾脏纤维化程度，抑制TGF-β1、Smad3、Smad4的基因和蛋白表达量，促进Smad7的表达量增多，可能通过调控TGF-β1/Smad信号通路而发挥相关治疗作用。

Abstract:

[Objective] To observe the effect of Erhuang Yishen Decoction(EHYST) on serum creatinine (BUN), serum creatinine (Scr), transforming growth factor-β1 (TGF-β1), Smad3, Smad4, Smad7 mRNA and protein expression levels in chronic renal failure via inactivation of the TGF-β1/Smad signal pathway.[Methods] Healthy SPF male SD rats were used to establish the animal model of chronic renal failure by 5/6 nephrectomy. After successful modeling, they were randomly divided into model group, Niaoduqing group(NDQ) and EHYST group. The drug groups were given the corresponding drugs for gavage treatment for 12 weeks. At the end of the experiment, BUN and Scr contents were measured, and pathological changes were analyzed by HE staining. TGF-β1, Smad3, Smad4 and Smad7 mRNA and protein expression were detected by RT-PCR and western blot.[Results] The animal model of chronic renal failure could be successfully established by 5/6 nephrectomy. Compared with the model group, the EHYST group and the NDQ group could reduce the Scr and BUN contents, and the difference was statistically significant(P<0.01). In the EHYST Decoction group and NDQ group, a small amount of inflammatory cells infiltrated and fibrous hyperplasia were observed in the renal interstitium, and the area of fibrosis was alleviated, which could improve the degree of renal fibrosis in renal failure. The results of gene and protein expression analysis showed that compared with the model group, the expression levels of TGF-β1, Smad3, and Smad4 in the EHYST Decoction group and NDQ group showed a decreasing trend, and the difference was statistically significant(P<0.05), and the expression levels of Smad7 showed an increasing trend, and the difference was also statistically significant(P<0.05).[Conclusion] EHYST could improve the degree of renal fibrosis, inhibit the gene and protein expression of TGF-β1, Smad3 and Smad4, and promote the increased expression of Smad7 in rats with chronic renal failure, which may play a related therapeutic role through regulating TGF-β1/Smad signaling pathway.