基于析因设计探讨六味地黄汤不同组方对5/6肾切除大鼠肾纤维化的影响

Investigate on the Effect of Different Formulations of Liuwei Dihuang Decoction on Renal Fibrosis in 5/6 Nephrectomy Rats Based on Factorial Design

目的基于析因设计探讨六味地黄汤不同组方对5/6肾切除大鼠术后的影响及机制。方法80只SD大鼠分为假手术组、模型组(A2B2)和六味地黄汤原方组(A1B1)及其拆方纯补组(A1B2)、纯泄组(A2B1)。模型组予0.9%生理盐水(NS)灌胃,其余组予以上述中药煎液灌胃。8周后检测大鼠的肾功能改变、残存肾病理改变,分别用免疫组化和RT-PCR法检测残肾组织Jaggedl、Notchl、α-SMA、E-cadherin蛋白和mRNA表达。结果大鼠5/6肾切除术后出现肾纤维化,肾功能下降,经六味地黄汤及其组方干预后,肾纤维化程度和肾功能均得到改善,残肾组织Notch1、Jaggedl、α-SMA蛋白和mRNA水平表达下降,E-cadherin蛋白和mRNA水平表达(P<0.05)。其中六味地黄汤原方组改善肾功能效果最明显。结论六味地黄汤“三补”“三泄”在改善大鼠5/6肾切除术后出现肾纤维化方面具有显著的协同增效效应,这可能与抑制残肾组织Notch1/Jaggedl信号通路激活,下调α-SMA和上调E-cadherin表达有关。

Objective Based on the factorial design to Investigate the effect and potential mechanism of Liuwei Dihuang Decoction in 5/6 nephrectomy rats.Methods The rats were divided into 5 groups:sham operation group,model group,Liuwei Dihuang Decoction group and " three tonic herbs" group and "three purgative herbs" group.The model group was gavage with 0.9%NS,while the rest groups were gavage with the above Chinese herbal decoction.After 8 weeks,we detected renal function changes and residual renal pathological changes in rats.We also detected the expressions of Jaggedl,Notchl,α-SMA,E-cadherin protein and mRNA respectively in rats’ remnant renal tissues by immunohistochemistry and RT-PCR.Results Renal fibrosis occured and renal function decreased in 5/6 nephrectomy rats.After the intervention of different formulations of Liuwei Dihuang decoction,degree of fibrosis and renal function both improved,and the expressions of Notchl,Jaggedl and SMA in remnant kidney tissues were decreased while the expressions of ECadherin increased. The improvement of liuwei Dihuang decoction group was the most obvious in different formulations of Liuwei Dihuang decoction groups.Conclusion The "three tonic herbs" and the "three purgative herbs" of Liuwei Dihuang decoction have significant synergistic effects in the improvement of renal fibrosis in 5/6 nephrectomy rats,which may be related to the inhibition of Notch 1/Jaggedl signaling pathway activation,down-regulation of-SMA and up-regulation of E-cadherin expression in the residual kidney.