抗纤灵二号方对单侧输尿管梗阻大鼠肾小管上皮细胞转分化的调节作用

Effects of "Kang Xian Ling Decoction-Ⅱ" on Renal Tubule Epithelial Mesenchymal Transdifferentiation in Rats After Unilateral Ureteral Obstruction

目的:通过研究中药复方——抗纤灵二号方对单侧输尿管梗阻(unilateral ureteral obstruction,UUO)大鼠肾间质α-平滑肌肌动蛋白(alpha-smooth muscle actin,α-SMA)、转化生长因子-β1(transforming growth factor-beta1,TGF-β1)、肝细胞生长因子(hepatocyte growth factor,HGF)表达的影响,探讨其对肾小管上皮细胞间充质转分化(epithelial mesenchymal transdifferentiation,EMT)的调节机制。方法:建立大鼠UUO模型,90只SD大鼠随机分为假手术组、模型组、代文组,及抗纤灵二号方低、中、高剂量组,于造模4周后腹主动脉采血检测血清肌酐(serum creatinine,Scr)、尿素氮(blood urea nitrogen,BUN),收集24h尿液检测N-乙酰-β-D-氨基葡萄糖苷酶(N-acetyl-β-D-glucosaminidase,NAG)、β2-微球蛋白(β2-mi-croglobulin,β2-MG),取梗阻侧肾组织,HE、Masson染色观察肾小管间质病变,免疫组织化学染色观察α-SMA、TGF-β1及HGF在肾间质的阳性染色表达,免疫印迹法检测肾组织α-SMA的蛋白表达,并进行半定量分析。结果:与假手术组相比,模型组大鼠Scr、BUN、尿NAG、β2-MG水平以及α-SMA、TGF-β1阳性表达显著升高(P<0.01)。与模型组相比,抗纤灵二号方高剂量组大鼠Scr、BUN水平降低,尿NAG、β2-MG排泄减少(P<0.01);抗纤灵二号方高剂量组和代文组α-SMA、TGF-β1阳性表达显著下调,而HGF阳性表达则显著上调(P<0.01),α-SMA蛋白表达也明显下调(P<0.05);低、中、高三个剂量组的疗效呈剂量依赖关系。结论:抗纤灵二号方可能通过改善肾小球、肾小管功能,抑制α-SMA、TGF-β1的表达,诱导HGF的高表达,从而抑制肾小管EMT,达到改善肾间质纤维化的作用。

Objective：To investigate the effects of ＂Kang Xian Ling Decoction- Ⅱ ＂（KXLDN- Ⅱ ）on the renal interstitium alpha- smooth muscle actin（α- SMA）, transforming growth factor - betal （TGF- β1 ）and bepatocyte growth factor（HGF）expression in unilateral ureteral obstruction（UUO）rats and to illuminate the possible mechanisms of renal tubule epithelial mesenchymal transdifferentiation（EMT）. Methods： 90 Sprague- Dawley rats were randomly divided into Shamoperated, model, Diovan, and （KXLDN-Ⅱ）treatment groups of low, middle and high dose. The rats were under intragastric administration of（KXLDN-Ⅱ ）after operation, and sodium chloride in tales doses in Sham and model groups. On the 4th week after treatment, detecting the serum creatinine（Scr）, blood urea nitrogen （BUN）, the N - acetyl - β - D - glucosaminidase（NAG） and β2 - microglobulin（ β2 - MG） in 24 h urine;the obstructed kidney was taken out to be measured by HE, Masson, and immunohistochemistry staining for semiquantitative analysis of α- SMA, TGF-β1 and HGF, and Western blotting for α- SMA. Results： The lever of Scr, BERN, NAG, β2- MG, α- SMA and TGF - β1 in model group increased as compared with those in the sham - operated group （ P 〈 0.01 ） In comparison with the model group, the lever of Scr, BUN, NAG and β2 - MG in KXLDN - Ⅱ treated groups of high dose were significantly lower （P 〈 0.01 ） ; and the expression of positive staining for a - SMA and TGF - β1 in KXLDN - Ⅱ treated groups of high dose and Diovan treated group were significantly down- regulated（P 〈 0.01） ;but the expression of HGF in KXLDN - Ⅱ treatment groups were significantly up - regulated（P 〈 0.01 ） ; the expression of α- SMA protein was down- regulated; the therapeutic effect of KXLDN- Ⅱ treated groups of low,middle and high dose rely on dose. Conclusion：KXLDN- Ⅱ may degrade renal interstitial fibrosis by improving renal function, inhibiting the expression of α - SMA and TGF - β1, inducing the expression of HGF, and inhibiting renal tubule EMT.