Central Laboratory, Seventh People's Hospital of Shanghai University of TCM, 358 Datong Road, Pudong, Shanghai, 200137, China.
Central Laboratory, Seventh People's Hospital of Shanghai University of TCM, 358 Datong Road, Pudong, Shanghai, 200137, China.
Biomed Pharmacother. 2017 Nov;95:520-528. doi: 10.1016/j.biopha.2017.08.115. Epub 2017 Sep 5.
The present study aimed to evaluate the protective effects of Erhuang Formula (EHF) and explore its pharmacological mechanisms on adenine-induced chronic renal failure (CRF).
The compounds in EHF were analyzed by HPLC/MS. Adenine-induced CRF rats were administrated by EHF. The effects were evaluated by renal function examination and histology staining. Immunostaining of some proteins related cell adhesion was performedin renal tissues, including E-cadherin, β-catenin, fibronectin and laminin. The qRT-PCR was carried out determination of gene expression related inflammation and fibrosis including NF-κB, TNF-α, TGF-β1, α-SMA and osteopontin (OPN).
Ten compounds in EHF were identified including liquiritigenin, farnesene, vaccarin, pachymic acid, cycloastragenol, astilbin, 3,5,6,7,8,3',4'-heptemthoxyflavone, physcion, emodin and curzerene. Abnormal renal function and histology had significant improvements by EHF treatment. The protein expression of β-catenin, fibronectin and laminin were significantly increased and the protein expression of E-cadherin significantly decreased in CRF groups. However, these protein expressions were restored to normal levels in EHF group. Furthermore, low expression of PPARγ and high expression of NF-κB, TNF-α, TGF-β1, α-SMA and OPN were substantially restored by EHF treatment in a dose-dependent manner.
EHF ameliorated renal damage in adenine-induced CRF rats, and the mechanisms might involve in the inhibition of inflammatory and fibrotic responses and the regulation of PPARγ, NF-κB and TGF-β signaling pathways.
本研究旨在评估二黄方（EHF）的保护作用，并探讨其对腺嘌呤诱导的慢性肾衰竭（CRF）的药理机制。
采用 HPLC/MS 分析 EHF 中的化合物。腺嘌呤诱导的 CRF 大鼠给予 EHF 治疗。通过肾功能检查和组织学染色评估疗效。免疫组化检测肾组织中与细胞黏附相关的某些蛋白，包括 E-钙黏蛋白、β-连环蛋白、纤连蛋白和层粘连蛋白。采用 qRT-PCR 法测定与炎症和纤维化相关的基因表达，包括 NF-κB、TNF-α、TGF-β1、α-SMA 和骨桥蛋白（OPN）。
鉴定出 EHF 中的 10 种化合物，包括甘草素、法呢烯、蝙蝠葛诺林碱、巴戟天甲素、环黄芪醇、毛蕊异黄酮、3,5,6,7,8,3',4'-七甲氧基黄酮、大黄素、大黄素甲醚和莪术烯。EHF 治疗可显著改善异常的肾功能和组织学变化。CRF 组β-连环蛋白、纤连蛋白和层粘连蛋白的蛋白表达显著增加，E-钙黏蛋白的蛋白表达显著降低；而 EHF 组这些蛋白表达恢复正常水平。此外，EHF 以剂量依赖性方式显著降低低表达的 PPARγ，同时显著升高高表达的 NF-κB、TNF-α、TGF-β1、α-SMA 和 OPN。
EHF 可改善腺嘌呤诱导的 CRF 大鼠的肾损伤，其机制可能涉及抑制炎症和纤维化反应以及调节 PPARγ、NF-κB 和 TGF-β 信号通路。
