“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-1α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

关键词:  肾间质纤维化  黄芪-丹参  单侧输尿管梗阻  缺氧诱导因子-1α

Key words:  renal interstitial fibrosis  astragalus-salvia  UUO  HIF-1α

摘要:

[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。

Abstract:

[Objective] To explore the effects of "astragalus-salvia" on renal interstitial fibrosis and renal tubular epithelial cell transdifferentiation (EMT) and the expression of HIF-1α through 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-1α protein expression;immunohistochemical method to detect rat α-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 of α-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-1α was 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-1α expression.