芪蓟肾康汤对转基因免疫球蛋白A肾病小鼠转化生长因子β1/Smads信号转导通路的调控机制研究

Regulatory mechanism of Qijishenkang Decoction on signaling pathway of transforming growth factorβ1/Smads in mice with gene immunoglobulin A nephropathy

目的观察芪蓟肾康汤对转基因免疫球蛋白A(IgA)肾病小鼠转化生长因子β1(TGF-β1)/Smads信号转导通路的影响,探讨其治疗IgA肾病的作用机制。方法将40只IgHA1-mut转基因F1代小鼠随机分为模型组、西药组、芪蓟肾康汤低剂量组、芪蓟肾康汤中剂量组及芪蓟肾康汤高剂量组,每组8只,另随机取8只C57BL/6J小鼠作为空白组。空白组及模型组按2 mL/100 g予0.9%氯化钠注射液灌胃,西药组按2 mg/100 g予替米沙坦药液灌胃,芪蓟肾康汤低、中、高剂量组分别按0.6、1.2、2.4 g生药/100 g灌胃,每日1次,连续灌胃28 d。比较2组灌胃14、21、28 d后肾组织Smad3 mRNA、Smad7 mRNA及TGF-β1、Smad 2蛋白表达情况。结果与空白组灌胃后同期比较,模型组灌胃14、21、28 d后Smad3 mRNA表达水平均明显升高(P<0.05),Smad7 mRNA表达水平均明显降低(P<0.05);与模型组灌胃后同期比较,西药组及芪蓟肾康汤中、高剂量组灌胃14、21、28 d后Smad3 mRNA表达水平均明显降低(P<0.05),Smad7 mRNA表达水平均明显升高(P<0.05),芪蓟肾康汤低剂量组仅灌胃28 d后Smad3 mRNA及Smad7 mRNA表达水平与模型组比较差异有统计学意义(P<0.05);芪蓟肾康汤各剂量组灌胃后组间同期比较,芪蓟肾康汤中、高剂量组灌胃14、21、28 d后Smad3 mRNA表达水平均明显低于芪蓟肾康汤低剂量组(P<0.05),Smad7 mRNA表达水平均明显高于芪蓟肾康汤低剂量组(P<0.05),且芪蓟肾康汤高剂量组灌胃14、21、28 d后Smad3 mRNA及Smad7 mRNA表达水平改善均优于同期芪蓟肾康汤中剂量组(P<0.05)。与空白组灌胃后同期比较,模型组灌胃14、21、28 d后TGF-β1及Smad2蛋白表达水平均明显升高(P<0.05);与模型组灌胃后同期比较,西药组及芪蓟肾康汤中、高剂量组灌胃14、21、28 d后TGF-β1及Smad2蛋白表达水平均明显降低(P<0.05),芪蓟肾康汤低剂量组仅灌胃21、28 d后TGF-β1及Smad2蛋白表达水平明显低于模型组(P<0.05);芪蓟肾康汤各剂量组灌胃后组间同期比较,芪蓟肾康汤中、高剂量组灌胃14、21、28 d后TGF-β1蛋白表达水平均明显低于芪蓟肾康汤低剂量组(P<0.05),且芪蓟肾康汤高剂量组灌胃28 d后TGF-β1蛋白表达水平均明显低于芪蓟肾康汤中剂量组(P<0.05)。结论芪蓟肾康汤对转基因IgA肾病小鼠治疗作用确切,可能是通过多个靶点调节TGF-β1/Smads信号转导通路完成,预防肾纤维化,延缓肾小球硬化过程。

Objective To observe the effects of Qijishenkang Decoction on the signal transduction pathway of TGF-β1/Smads in mice with transgenic immunoglobulin A(IgA)nephropathy,and to explore the treatment mechanism of Qijishenkang Decoction on IgA nephropathy.Methods Forty IgHA1-mut transgenic F1 generation of mice were divided into the model group,western medicine group,Qijishenkang Decoction low-dose group,Qijishenkang Decoction medium-dose group and Qijishenkang Decoction high-dose group with 8 mice in each group,and another 8 C57BL/6J mice were divided into the blank group.The mice in the blank group and model group were intragastrically administrated with 0.9%Sodium Chloride Injection at the dose of 2 ml/100 g,the mice in the western medicine group was administered with telmisartan by gavage at the dose of 2 mg/100 g,and the mice in the low-dose,medium-dose and high-dose Qijishenkang Decoction groups were administered with 0.6,1.2 and 2.4 g/100 g of crude drugs by gavage once a day for 28 consecutive days.The expressions of Smad3 mRNA,Smad7 mRNA,TGF-β1 protein and Smad2 protein in renal tissue between two groups at 14,21 and 28 days after intragastric administration were compared.Result Compared to the indexes during the same period after intragastric administration in the normal group,the expression level of Smad3 mRNA in mice in the model group was significantly elevated(P<0.05),and the expression level of Smad7 mRNA was significantly lowered(P<0.05)in the 14th,21st and 28th day after intragastric administration;Compared to the index during the same period after intragastric administration in the model group,the expression level of Smad3 mRNA in the western medicine group,Qijishenkang Decoction medium-dose and high-dose groups was significantly lowered(P<0.05),and the expression level of Smad7 mRNA was significantly lowered(P<0.05);The expression levels of Smad3 mRNA and Smad7 mRNA of mice in low-dose Qijishenkang Decoction group were significantly higher than such indexes of mice in the model group(P<0.05),expression levels of Smad3 mRNA and Smad7 mRNA in the medium-dose and high-dose Qijishenkang Decoction groups were significantly higher than those in the model group(P<0.05),the expression level of Smad7 mRNA was higher than that of low-dose Qijishenkang Decoction group(P<0.05),and the expression level of Smad3 mRNA and Smad7 mRNA of high-dose Qijishenkang Decoction group was better than such index of mice in medium-dose Qijishenkang Decoction group(P<0.05).Compared to the indexes of mice during the same period after intragastric administration in the normal group,protein expression levels of TGF-β1 and Smad2 of mice in the model group were significantly increased in the 14th,21st and 28th days after intragastric administration(P<0.05);Compared to the indexes during the same period after intragastric administration in the model group,the protein expression levels of TGF-β1 and Smad2 of mice in the western medicine group and medium-dose and high-dose Qijishenkang Decoction groups were significantly decreased in the 14th,21st and 28th days after intragastric administration(P<0.05),and the protein expression levels of TGF-β1 and Smad2 of mice in low-dose Qijishenkang Decoction administered on the 21st and 28th day after intragastric administration were significantly lower than such indexes of mice in the model group(P<0.05);The protein expression level of TGF-β1 in medium-dose and high-dose Qijishenkang Decoction groups in the 14th,21st and 28th days after intragastric administration were significantly lower than such indexes of mice in the low-dose Qijishenkang Decoction group(P<0.05),and the expression level of TGF-β1 protein of mice in high-dose Qijishenkang Decoction group in the 28th day after intragastric administration was significantly lower than such index of mice in Qijishenkang Decoction low-dose group and Qijishenkang Decoction medium-dose group(P<0.05).Conclusion:Qijishenkang Decoction delivers definite therapeutic effects on transgenic IgA nephropathy mice,and the effects can be achieved by regulating TGF-β1/Smads signal transduction pathway through multiple targets,delaying the process of glomerulosclerosis.