“黄芪-丹参”对单侧输尿管梗阻大鼠肾间质纤维化及低氧诱导因子-1 α表达的影响

Effect of “astragalus-salvia” on renal interstitial fibrosis and hypoxia inducible Factor-1α expression in rats with unilateral ureteral obstruction

[目的]通过单侧输尿管梗阻(UUO)大鼠模型探讨“黄芪-丹参”对于肾间质纤维化和肾小管上皮细胞转分化(EMT)的作用以及缺氧诱导因子(HIF)-1α表达的影响。[方法]将大鼠分为假手术组、模型组、芪参低剂量组、芪参中剂量组、芪参高剂量组、氯沙坦组。采用UUO模型并灌胃、取材。Masson染色法观察大鼠肾脏组织病理改变;免疫组化法检测大鼠肾脏组织α-平滑肌肌动蛋白(α-SMA)表达;PCR法检测大鼠肾脏组织1型胶原蛋白(Col1)、纤连蛋白(FN)mRNA表达;蛋白免疫印迹(Western Blot)法检测大鼠肾脏组织HIF-1α蛋白表达。[结果]Masson染色:模型组与假手术组比较,胶原容积升高,差异具有统计学意义(P<0.001);芪参中、高剂量与模型组比较出现下降,差异具有统计学意义(P<0.01);氯沙坦组与模型组比较下降,差异具有统计学意义(P<0.001)。免疫组化:大鼠模型组与假手术组比较,α-SMA蛋白表达升高,差异具有统计学意义(P<0.001);芪参中、高剂量以及氯沙坦组与模型组比较,α-SMA均下降,差异具有统计学意义(P<0.01)。实时荧光定量聚合酶链式反应(PCR)检测:模型组对比假手术组,Col1、FN表达水平升高(P<0.001),药物组与模型组比较Col1、FN表达降低,差异具有统计学意义(P<0.05)。Western Blot检测:模型组与假手术组比较,HIF-1α表达升高,差异具有统计学意义(P<0.001);芪参中剂量较模型组有下降,差异具有统计学意义(P<0.01)。[结论]“黄芪-丹参”药对可以通过抑制细胞外基质的生成以及减轻肾小管上皮细胞-间充质细胞转分化两个方面延缓肾间质纤维化,其机制可能与抑制HIF-1α表达有关。

[Objective]To explore the effects of“astragalus-salvia”on renal interstitial fibrosis and renal tubular epithelial cell transdifferentiation(EMT)and the expression of HIF-1αthrough the unilateral ureteral obstruction(UUO)model.[Methods]The rats were divided into sham operation group,model group,low-dose Chinese medicine group,middle-dose Chinese medicine group,high-dose Chinese medicine group,and Losartan group.The UUO model was used and the stomach was given and the materials were taken.Observe rat Masson staining;Western Blot method to detect HIF-1αprotein expression;immunohistochemical method to detect ratα-SMA protein expression;PCR to detect rat Col1 and FN mRNA expression.[Results]Masson staining:compared with the model group and the sham operation group,the collagen volume was significantly increased(P<0.001);the medium and high doses of Chinese medicine decreased compared with the model group(P<0.01);Losartan group was significantly lower than model group(P<0.001).Immunohistochemistry:compared with the sham operation group,the expression ofα-SMA protein increased significantly in the rat model group(P<0.001);compared with the model group in the middle and high doses of Chinese medicine and the losartan group,α-SMA decreased significantly(P<0.01).Real-time fluorescence quantitative PCR detection:compared with the sham operation group,the expression levels of Col1 and FN increased significantly in the model group(P<0.001),and the expression levels of Col1 and FN decreased in the drug group compared with the model group(P<0.05).Western Blot detection:compared with the sham operation group,the expression of HIF-1αwas significantly increased in the model group(P<0.001);the dose of Chinese medicine was significantly lower than that in the model group(P<0.01).[Conclusion]“Astragalus-salvia”can delay renal interstitial fibrosis by inhibiting the production of extracellular matrix and reducing the transdifferentiation of renal tubular epithelial cells-mesenchymal cells.The mechanism may be related to the inhibition of HIF-1αexpression.