Li Aiping, Wang Mengjiao, Guo Min, Liu Yuetao, Li Ke, Zhang Lichao, Qin Xuemei, Liu Guangzhen
Modern Research Center for Traditional Chinese Medicine of Shanxi University, Taiyuan, Shanxi 030006, China; Shanxi Academy of Traditional Chinese Medicine, Taiyuan 030012, China.
Modern Research Center for Traditional Chinese Medicine of Shanxi University, Taiyuan, Shanxi 030006, China.
Mol Immunol. 2025 Oct;186:161-173. doi: 10.1016/j.molimm.2025.08.015. Epub 2025 Aug 25.
Astragali Radix (AR), a homologous of medicine and food, has been extensively recorded to possess a nephroprotective impact on individuals suffering from chronic kidney disease (CKD). Astragaloside IV (ASIV) is one of the prominent bioactive constituents derived from AR. This study aimed to investigate how ASIV promotes M2 polarization of macrophages and whether this contributes to the protection of podocytes from injury, using a combination of lipidomics and molecular biology techniques.The effect of ASIV on adriamycin (ADR)-induced renal injury in rats was evaluated in vivo, with a particular emphasis on elucidating the potential involvement of macrophages. The M1 polarization model of RAW264.7 cells induced by ADR was established in vitro. The impact of ASIV on ADR-induced macrophage polarization was comprehensively assessed by measuring the expression levels of M1 and M2 macrophage marker proteins, along with their associated mRNA profiles. Flow cytometry was employed to analyze surface marker expression, while Western blot (WB) was used to quantify protein levels, and real-time quantitative PCR (qPCR) allowed for the measurement of gene expression. Notably, we found that the macrophage supernatant intervened by ASIV was found to have a protective effect against podocyte injury, which was evaluated through cell adhesion and apoptosis assays. Innovatively, we explored the mechanism by which ASIV affects macrophage polarization from the perspective of lipid metabolism, using lipidomics methods. During this process, the role of peroxisome proliferator-activated receptor gamma (PPARγ) in macrophage polarization caught our attention. This receptor is closely associated with lipid metabolites, as confirmed by molecular docking. ASIV has been shown to effectively ameliorate kidney injury, with macrophages playing a pivotal role in renal podocyte repair process. The protein and mRNA expressions of ARG-1, CD206 and IL-10, M2 macrophage markers, was increased by ASIV and M2 macrophages polarization was promoted by ASIV (P < 0.05). In addition, it was observed that the supernatant of macrophages intervened by ASIV exerts a protective effect on podocyte injury, which was confirmed through podocyte adhesion assays and cell apoptosis experiments. This study suggests that ASIV enhances the M2 polarization of macrophages, and the supernatant from these polarized macrophages has a protective effect on podocytes. From the perspective of lipid metabolism, the underlying mechanism of this effect may involve the modulation of sphingolipids, arachidonic acid, glycerophospholipids, and other lipid metabolites, which activate the receptor PPARγ, thereby exerting protective effects on podocytes.
黄芪，一种药食同源的物质，已有大量记载表明其对慢性肾脏病（CKD）患者具有肾脏保护作用。黄芪甲苷（ASIV）是从黄芪中提取的一种重要生物活性成分。本研究旨在结合脂质组学和分子生物学技术，探讨ASIV如何促进巨噬细胞的M2极化以及这是否有助于保护足细胞免受损伤。在体内评估了ASIV对阿霉素（ADR）诱导的大鼠肾损伤的影响，特别着重于阐明巨噬细胞的潜在作用。在体外建立了由ADR诱导的RAW264.7细胞M1极化模型。通过测量M1和M2巨噬细胞标志物蛋白的表达水平及其相关的mRNA谱，全面评估了ASIV对ADR诱导的巨噬细胞极化的影响。采用流式细胞术分析表面标志物表达，蛋白质印迹法（WB）定量蛋白质水平，实时定量聚合酶链反应（qPCR）测量基因表达。值得注意的是，我们发现经ASIV干预的巨噬细胞上清液对足细胞损伤具有保护作用，这通过细胞黏附和凋亡试验得以评估。创新性地，我们利用脂质组学方法从脂质代谢的角度探讨了ASIV影响巨噬细胞极化的机制。在此过程中，过氧化物酶体增殖物激活受体γ（PPARγ）在巨噬细胞极化中的作用引起了我们的关注。经分子对接证实，该受体与脂质代谢产物密切相关。已证明ASIV能有效改善肾损伤，巨噬细胞在肾足细胞修复过程中起关键作用。ASIV增加了M2巨噬细胞标志物ARG-1、CD206和IL-10的蛋白质和mRNA表达，并促进了M2巨噬细胞极化（P<0.05）。此外，观察到经ASIV干预的巨噬细胞上清液对足细胞损伤具有保护作用，这通过足细胞黏附试验和细胞凋亡实验得到证实。本研究表明，ASIV增强了巨噬细胞的M2极化，这些极化巨噬细胞的上清液对足细胞具有保护作用。从脂质代谢的角度来看，这种作用的潜在机制可能涉及鞘脂、花生四烯酸、甘油磷脂和其他脂质代谢产物的调节，这些物质激活了PPARγ受体，从而对足细胞发挥保护作用。
