State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College and Beijing Key Laboratory of Drug Target and Screening Research, Beijing, 100050, China.
Faculty of Biology, Medicine and Health Sciences, The University of Manchester Oxford Road, Manchester, M13 9PL, United Kingdom.
J Ethnopharmacol. 2024 Jan 30;319(Pt 2):117281. doi: 10.1016/j.jep.2023.117281. Epub 2023 Oct 4.
Puerarin (PUR) isolated from the root of Pueraria lobata (Willd.) Ohwi is considered as one of the main medicines to alleviate asthenic splenonephro-yang of diabetic nephropathy (DN). Whereas, the exact mechanism of Puerarin on diabetic nephropathy is still unclear.
In this study, we aimed to investigate the protective effects of PUR on type 2 diabetic nephropathy in vivo, in silico and in vitro, as well as unveil the underlying mechanism through inhibiting ferroptosis.
In vivo, blood glucose and lipid, renal function, kidney histology and immunohistochemistry analysis were used to vindicate the protective effects of PUR on diabetic nephropathy in type 2 DN rat model. In silico, pharmacophore matching and enrichment analysis were adopted to predict the potential mechanism of PUR on DN. In vitro, we utilized high glucose stress to induce impairment in glomerular mesangial cells (GMCs) as diabetic nephropathy cell model. Cell count kit-8 (CCK-8) was used to observe cell viability. qPCR, Western blot, immunofluorescence staining and flow cytometry were used to evaluate the effect of PUR on the generation of extracellular matrix (ECM), ferroptosis and iron homeostasis in vitro and in vivo.
PUR markedly improved glucose and lipid metabolism, as well as alleviated renal dysfunction in diabetic nephropathy rats. Pharmacophore matching and enrichment analysis predicted the anti-DN effect of PUR may correlate with ECM. Experimental validation suggested that PUR treatment could inhibit the generation of ECM to alleviate high-glucose-induced cell impairments, suppressing ROS production and excessive collagen fiber accumulation in GMSs, and reduce mesangial matrix expansion and renal fibrosis in type 2 DN rats. Further study suggested that PUR protected GMCs against ferroptosis via reducing LDH release and GSH disruption, suppressing key regulators of two pathways for ferroptosis execution. Moreover, PUR also maintained iron metabolism hemostasis by regulating iron transportation proteins, iron exporter proteins, and iron storage proteins and reducing intracellular iron in type 2 DN rats.
PUR inhibited excessive ECM accumulation to protect against type 2 diabetic nephropathy, which meditated by regulating iron homeostasis and mitigating ferroptosis. This study provides promising therapeutics for diabetic nephropathy treatment.
葛根素（PUR）从野葛（Willd.）Ohwi 的根部中分离出来，被认为是缓解糖尿病肾病（DN）虚脾肾虚的主要药物之一。然而，葛根素治疗糖尿病肾病的确切机制尚不清楚。
本研究旨在通过体内、体外和计算方法研究 PUR 对 2 型糖尿病肾病的保护作用，并通过抑制铁死亡来揭示其潜在机制。
在体内，我们使用血糖和血脂、肾功能、肾脏组织学和免疫组织化学分析来验证 PUR 对 2 型糖尿病肾病大鼠模型的保护作用。在计算方面，我们采用药效团匹配和富集分析来预测 PUR 对 DN 的潜在作用机制。在体外，我们利用高糖应激诱导肾小球系膜细胞（GMCs）损伤作为糖尿病肾病细胞模型。细胞计数试剂盒-8（CCK-8）用于观察细胞活力。qPCR、Western blot、免疫荧光染色和流式细胞术用于评估 PUR 对体外和体内细胞外基质（ECM）生成、铁死亡和铁平衡的影响。
PUR 显著改善了糖尿病肾病大鼠的糖脂代谢，改善了肾功能。药效团匹配和富集分析预测 PUR 的抗 DN 作用可能与 ECM 相关。实验验证表明，PUR 治疗可抑制 ECM 的生成，从而缓解高糖诱导的细胞损伤，抑制 GMSs 中 ROS 产生和胶原纤维过度积累，减少系膜基质扩张和 2 型糖尿病大鼠的肾纤维化。进一步的研究表明，PUR 通过减少 LDH 释放和 GSH 破坏、抑制两条铁死亡途径的关键调节因子来保护 GMCs 免受铁死亡。此外，PUR 还通过调节铁转运蛋白、铁输出蛋白和铁储存蛋白以及减少 2 型糖尿病大鼠细胞内铁来维持铁代谢平衡。
PUR 通过调节铁平衡和减轻铁死亡来抑制 ECM 的过度积累，从而防止 2 型糖尿病肾病，为糖尿病肾病的治疗提供了有前途的治疗方法。
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