[关键词]
[摘要]
目的 观察温阳泄浊通络法对5/6肾切除所致慢性肾衰竭大鼠肾间质纤维化的干预作用,并探讨其机制。方法 32只SD大鼠随机选取8只为假手术组,其余24只行5/6肾切除术制备肾纤维化模型。造模完成后随机分为模型组、温阳泄浊通络法组、尿毒清组,并ig予以相应药物。干预12周后,取肾组织,行HE、PAS染色检测病理损伤程度,行Masson、天狼星红染色检测纤维化程度,并应用免疫荧光、Western blotting法检测I型胶原蛋白及mRNA的表达,应用免疫组化、Western blotting法检测肾小管上皮细胞EMT相关蛋白(α-SMA、Vimentin、E-cadherin)的表达。结果 与模型组比较,温阳泄浊通络法可减轻肾脏病理损伤及肾纤维化程度,并显著降低I型胶原蛋白及mRNA的表达;下调α-SMA、Vimentin的高表达,上调E-cadherin的低表达。结论 温阳泄浊通络法可有效减轻慢性肾衰竭大鼠肾损伤、改善肾纤维化,其机制可能与抑制肾小管上皮细胞EMT及I型胶原的表达相关。
[Key word]
[Abstract]
Objective To investigate the effects of warming yang-dispelling turbidity-dredging collaterals method on renal interstitial fibrosis in rats with chronic renal failure caused by 5/6 nephrectomy and explore its mechanisms. Methods Eight rats were randomly chosen as sham operation group (Sham), while the remaining 24 rats underwent 5/6 nephrectomy were divided into model group (5/6Nx), warming yang-dispelling turbidity-dredging collaterals method group (WDD) and Niaoduqing group (NDQ), and the rats in each group were administrated with corresponding drugs. After the intervention of 12 weeks, all the rats were sacrificed and the kidneys were then removed. HE, PAS Masson, Picrosirius red staining were conducted to assess kidney pathological changes and renal fibrosis. The protein expression of Collagen Ⅰ was tested by both immunofluorescence and Western blotting, and the mRNA expression was evaluated using qRT-PCR. The expression of protein related with the process of EMT, including α-SMA, E-Cadherin and Vimentin, were tested with immunohistochemistry and Western blotting. Results Compared with the model group, warming yang-dispelling method alleviated kidney pathological injury and renal fibrosis, and significantly down-regulated the protein and mRNA expression of Collagen Ⅰ. Moreover, warming yang-dispelling turbidity-dredging collaterals method regulated the process of EMT by decreasing the high expression of α-SMA and Vimentin, and increasing the low expression of E-Cadherin. Conclusion Warming yang-dispelling turbidity-dredging collaterals method can effectively attenuate renal fibrosis and injury and its underlying mechanisms may be related with its inhibitory effects on Collagen Ⅰ expression and regulation of EMT.
[中图分类号]
R285.5
[基金项目]
河北省科技支撑计划项目(14277708D);河北省研究生创新资助项目(CXZZBS2018158)