Hunan Key Laboratory of Kidney Disease and Blood Purification, Department of Nephrology, The Second Xiangya Hospital, Central South University, Changsha, People's Republic of China.
Drug Des Devel Ther. 2023 Mar 23;17:887-900. doi: 10.2147/DDDT.S402503. eCollection 2023.
Accumulating clinical evidence showed that (Lév.) Hutch (THH) is effective against IgA nephropathy (IgAN), but the mechanism is still unclear. This study is to evaluate the renal protective effect and molecular mechanism of THH against IgAN via network pharmacology, molecular docking strategy and experimental validation.
Several databases were used for obtaining the active ingredients of THH, the corresponding targets, as well as the IgAN-related genes. The critical active ingredients, functional pathways, and potential for the combination of the hub genes and their corresponding active components were determined through bioinformatics analysis and molecular docking. The IgAN mouse model was treated with celastrol (1 mg/kg/d) for 21 days, and the aggregated IgA1-induced human mesangial cell (HMC) was treated with various concentrations of celastrol (25, 50 or 75 nM) for 48 h. The immunohistochemistry and Western blot techniques were applied to evaluate the protein expression of the predicted target. The cell counting kit 8 (CCK8) was used to detect HMC proliferation.
A total of 17 active ingredients from THH were screened, covering 165 IgAN-related targets. The PPI network identified ten hub targets, including PTEN. The binding affinity between the celastrol and PTEN was the highest (-8.69 kJ/mol). The immunohistochemistry showed that celastrol promoted the expression of PTEN in the glomerulus of IgAN mice. Furthermore, the Western blot techniques showed that celastrol significantly elevated the expression of PTEN and inhibited PCNA and Cyclin D1 in vitro and in vivo. The CCK8 assay determined that celastrol decreased HMC proliferation in a concentration-dependent manner.
This study suggests that activating PTEN by celastrol may play a pivotal role in THH alleviating IgAN renal injury.
越来越多的临床证据表明，（Lev.） Hutch（THH）对 IgA 肾病（IgAN）有效，但作用机制尚不清楚。本研究通过网络药理学、分子对接策略和实验验证，评估 THH 对 IgAN 的肾脏保护作用及分子机制。
利用多个数据库获取 THH 的活性成分、相应靶点以及 IgAN 相关基因。通过生物信息学分析和分子对接，确定关键活性成分、功能通路以及关键基因与相应活性成分结合的潜力。采用雷公藤红素（1mg/kg/d）处理 IgAN 小鼠模型 21 天，用不同浓度的雷公藤红素（25、50 或 75nM）处理人肾小球系膜细胞（HMC）48h。免疫组化和 Western blot 技术用于评估预测靶点的蛋白表达。细胞计数试剂盒 8（CCK8）检测 HMC 增殖。
从 THH 中筛选出 17 种活性成分，涵盖 165 个 IgAN 相关靶点。PPI 网络鉴定出 10 个关键靶点，包括 PTEN。雷公藤红素与 PTEN 的结合亲和力最高（-8.69kJ/mol）。免疫组化显示雷公藤红素促进 IgAN 小鼠肾小球中 PTEN 的表达。此外，Western blot 技术表明雷公藤红素在体外和体内均显著上调 PTEN 的表达，并抑制 PCNA 和 Cyclin D1 的表达。CCK8 测定表明雷公藤红素浓度依赖性地降低 HMC 增殖。
本研究表明，雷公藤红素通过激活 PTEN 可能在 THH 缓解 IgAN 肾损伤中发挥关键作用。
