归经组方对肾纤维化大鼠肾组织中Wnt信号表达的影响

Effect of Composing Prescription with Meridian Tropism on Wnt Signal Pathway in Renal Fibrosis Rats

目的:观察归肾经药物组方对肾纤维化大鼠疗效及肾组织Wnt信号表达的影响。方法:用5/6肾切除方法建立大鼠肾纤维化模型。经验方肾衰Ⅰ号为肾衰Ⅰ号组,以治则相同不归肾经药物组方为不归经组,以治则相同且归肾经的组方为归经组,同时设模型组,假手术组。连续治疗2月后,采血检测肾功能及血色素、取肾组织Masson染色、Western blot法检测肾组织Wnt2、GSK-3β、P-GSK-3β、β-catenin、α-SMA、β-actin蛋白表达,免疫组化法检测转化生长因子-β1(transforming growth factor-β1,TGF-β1)在肾组织表达。结果:成功造模并治疗2个月后,与假手术组比较,模型组体质量、血浆总蛋白(total protein,TP)、白蛋白(Albumin,Alb)、血红蛋白(hemoglobin,Hb)含量显著降低(P<0.01),血尿素氮(blood urea nitrogen,BUN)、血肌酐(creatinine,Cr)含量显著增高(P<0.01),Wnt2、P-GSK-3β、β-catenin、α-SMA及TGF-β1蛋白表达显著增高(P=0.000),GSK-3β活性显著降低(P=0.000),胶原含量显著增高(Masson染色)(P<0.05);与模型组比较,各治疗组体质量显著增高(P<0.01),BUN、Cr显著降低(P<0.01),Wnt2、P-GSK-3β、β-catenin、α-SMA、TGF-β1表达及胶原含量显著降低(P<0.05)。3个治疗组间相比较,归经组Hb、Tp含量及GSK-3β表达较肾衰Ⅰ号组及不归经组显著升高P<0.01),P-GSK-3β表达显著降低(P<0.01);不归经组α-SMA、β-catenin表达明显高于其他两组(P<0.05)。3个治疗组间体质量、ALB、BUN、Cr含量及Wnt2表达之间没有明显差异,归经组与肾衰Ⅰ号之间α-SMA、β-catenin表达亦没有明显差异。结论:三组方均能通过Wnt信号通路减轻模型动物肾纤维化程度,且经验方与归肾经处方优于不归肾经方。

Objective： To investigate the clinical efficacy the prescription composed with Kidney meridian tropism on renal fibrosis rats and the effect on the Wnt signal pathway. Method： The rat model of renal fibrosis was established with 5/6 nephrectomy. The empirical prescription was used in the first renal failure group; the composed prescription by using different kinds of drugs without Kidney meridian tropism was used in the second renal failure group; the composed prescription by using different kinds of drugs of Kidney meridian tropism was used in the third renal failure group. The forth group was model group and the fifth group was sham operation group. The expression of Wnt2,GSK-3β,P-GSK-3β,β-catenin,α-SMA and β-actin protein in renal tissue were detected by using Western blot and the expression of Wnt2,GSK-3β The expression of transforming growth factor-β1（ TGF-β1） were detected by using immunohistochemistry in renal tissue. Results： After 2 months of modeling and treatment,the contents of total protein（ TP）,albumin（ Alb）,hemoglobin（ Hb） were significantly decreased in the model group compared with that of the sham operation group（ P〈0. 01）. The levels of blood urea nitrogen（ BUN） and creatinine（ Cr） significantly increased（ P〈0. 01）,Wnt2,P-GSK-3β,β-catenin,α-SMA and TGF（ P = 0. 000）,GSK-3β activity significantly decreased（ P = 0. 000）,and collagen content significantly increased（ Masson staining）（ P〈0. 05）. Compared with that of the model group,the body weight of each treatment group was significantly higher（ P〈0. 01）,BUN and Cr significantly decreased（ P〈0. 01）,Wnt2,P-GSK-3β,β-catenin,α-SMA,TGF-β1 and collagen content significantly decreased（ P〈0. 05）. The expression of P-GSK-3β significantly decreased in the treatment group（ P〈0. 01）,and the expression of Hb,Tp and GSK-3β in the group was significantly higher than that in the first group and the second group（ P〈0. 01）. The expression of α-SMA and β-catenin in the second group was significantly higher than that in the other two groups（ P〈0. 05）. There was no significant difference in the contents of ALB,BUN,Cr and Wnt2 between the three treatment groups. There was no significant difference in the expression of α-SMA and β-catenin between the group and the renal failure group. Conclusion： The three groups can reduce the degree of renal fibrosis by Wnt signaling pathway,and the clinical efficacy of the empirical prescription and the prescription composed by using different kinds of drugs with Kidney meridian tropism are significantly better than other pricriptions.