温阳消癥方对5/6肾切除小鼠肾纤维化的作用及其机制

目的：观察温阳消癥方对5/6肾切除小鼠残肾功能及肾脏纤维化的影响，并探讨其作用机制。方法：随机从32只小鼠中取24只制作5/6肾切除慢性肾功能衰竭肾纤维化模型，2周后根据血清肌酐值将24只小鼠分为5/6肾切除组、代文组和温阳消癥方组，另8只作为假手术组。治疗8周后，Western blot检测各组小鼠肾组织JAK2、STAT3、p-JAK2、p-STAT3、转化生长因子β1（TGF-β1）、Smad3、I型胶原蛋白（Col-I）、III型胶原蛋白（Col-III）、α-SMA蛋白表达水平，免疫组织化学染色法测定小鼠肾组织Col-I、Col-III、α-SMA水平，用RT-qPCR法测定TGF-β1、Smad3 mRNA水平。结果：治疗8周后，模型组小鼠肾组织JAK2、STAT3、p-JAK2、p-STAT3、TGF-β1、Smad3、Col-I、Col-III、α-SMA蛋白表达均较假手术组上调（P＜0.01），模型组小鼠肾组织TGF-β1、Smad3 mRNA表达较假手术组上调（P＜0.01）；与模型组比较，代文组、温阳消癥方组JAK2、STAT3、p-JAK2、p-STAT3、TGF-β1、Smad3、Col-I、Col-III、α-SMA蛋白表达和TGF-β1、Smad3 mRNA表达均下调（P＜0.01）；与代文组比较，除TGF-β1、α-SMA外，温阳消癥方组其余指标均下调（P＜0.01）。结论：温阳消癥方能够保护5/6肾切除小鼠残肾功能，减轻肾纤维化病变程度，其机制可能与抑制TGF-β1/Smad3、JAK2/STAT3通路相关。

Effect of Wenyangxiaozheng decoction on renal fibrosis in 5/6 nephrectomy mice and its possible mechanism

Objective: To observe the effect of Wenyangxiaozheng decoction on renal function and renal fibrosis in 5/6 nephrectomy (5/6N) mice and explore its mechanism. Methods: Out of 32 C57 mice 24 were randomly chosen to establish 5/6N model which closely simulated the process of human chronic renal failure and renal fibrosis. Two weeks after the operation, 24 mice were divided as 5/6N group, diovan group and wenyangxiaozheng decoction group according to the value of serum creatinine. The other 8 mice were treated as sham operation group. Eight weeks after the treatment, the protein expression of JAK2, STAT3, p-JAK2, p-STAT3, transforming growth factor-β1 (TGF-β1), Smad3 was determined by Western blot; the protein expression of Collagen-I (Col-I), Collgaen-III (Col-III) and α-SMA was determined by both Western blot and immunohistochemistry, TGF-β1, Smad3 mRNA were determined by RT-qPCR. Results: Eight weeks after the treatment, the protein expression of JAK2, STAT3, p-JAK2, p-STAT3, TGF-β1, Smad3, Col-I, Col-III, α-SMA and TGF-β1, Smad3 mRNA expression in 5/6N group was significantly upregulated when compared with that of sham group (P<0.01); compared with 5/6N group, the protein expression of JAK2, STAT3, p-JAK2, p-STAT3, TGF-β1, Col-I, Col-III, α-SMA and TGF-β1, Smad3 mRNA expression was obviously downregulated in diovan group and wenyangxiaozheng decoction group (P<0.01); compared with diovan group, indexes were all downregulated in wenyangxiaozheng decoction group (P<0.01) except TGF-β1 and α-SMA. Conclusion: Wenyangxiaozheng decoction could protect renal function and reduce the degree of renal fibrosis in 5/6N mice, and the mechanism may be related to the inhibition of TGF-β1/Smad3 and JAK2/STAT3 signaling pathway.