Hyodo Mizusa, Kawada Kei, Ishida Tomoaki, Izawa-Ishizawa Yuki, Matoba Ryoko, Okamoto Rina, Jobu Kohei, Horikawa Io, Aizawa Fuka, Yagi Kenta, Niimura Takahiro, Kawano Yayoi, Abe Shinji, Hamada Yukihiro, Goda Mitsuhiro, Ishizawa Keisuke
Department of Clinical Pharmacology and Therapeutics, Tokushima University Graduate School of Biomedical Sciences, 3-18-15, Kuramoto-cho, Tokushima 770-8503, Japan.
Department of Pharmacy, Tokushima University Hospital, 2 Chome-50-1 Kuramoto-cho, Tokushima 770-8503, Japan.
Pharmaceuticals (Basel). 2025 Jul 24;18(8):1099. doi: 10.3390/ph18081099.
(Thunb.) DC. [Asteraceae] (ALR)-derived exosome-like nanoparticles (ALR-ELNs) exhibit anti-neuroinflammatory effects in microglial cells. However, the associated mechanisms and pathways are unknown. We aimed to characterize the effects of ALR-ELNs on inflammatory responses of BV-2 microglial cells to lipopolysaccharide (LPS) using RNA sequencing. ALR-ELNs were fractionated from ALR. BV-2 microglial murine cells were stimulated with LPS after treatment with ALR-ELNs. RNA sequencing was performed to analyze variations in mRNA levels. Ingenuity pathway analysis (IPA) was performed to investigate the mechanism of action of ALR-ELNs. mRNA expression was assessed using real-time quantitative polymerase chain reaction (qPCR). The expression of 651 genes was downregulated, whereas that of 1204 genes was upregulated in LPS-stimulated BV2 cells pretreated with ALR-ELNs. The IPA showed that the effects of ALR-ELNs on inflammation took place through pathogen-influenced signaling. Network analysis via IPA showed that the Toll-like receptor (TLR) is involved in the suppression of inflammation by ALR-ELNs. The qPCR analysis showed that pretreatment with ALR-ELNs significantly reduced mRNA expression. ALR-ELNs suppress the release of inflammatory mediators by downregulating expression, which is a novel mechanism by which ALR-ELNs act on microglia. Identifying active ingredients in ALR-ELNs that downregulate expression can advance the development of therapeutic drugs for neuroinflammatory diseases.
菊科植物苍耳（(Thunb.) DC. [Asteraceae]）衍生的类外泌体纳米颗粒（ALR-ELNs）在小胶质细胞中表现出抗神经炎症作用。然而，其相关机制和途径尚不清楚。我们旨在通过RNA测序来表征ALR-ELNs对BV-2小胶质细胞对脂多糖（LPS）炎症反应的影响。从苍耳中分离出ALR-ELNs。用ALR-ELNs处理BV-2小胶质鼠细胞后，再用LPS刺激。进行RNA测序以分析mRNA水平的变化。进行 Ingenuity 通路分析（IPA）以研究ALR-ELNs的作用机制。使用实时定量聚合酶链反应（qPCR）评估mRNA表达。在用ALR-ELNs预处理的LPS刺激的BV2细胞中，651个基因的表达下调，而1204个基因的表达上调。IPA表明，ALR-ELNs对炎症的影响是通过病原体影响的信号传导发生的。通过IPA进行的网络分析表明，Toll样受体（TLR）参与了ALR-ELNs对炎症的抑制作用。qPCR分析表明，用ALR-ELNs预处理可显著降低mRNA表达。ALR-ELNs通过下调[此处原文可能缺失某个基因或蛋白名称]的表达来抑制炎症介质的释放，这是ALR-ELNs作用于小胶质细胞的一种新机制。鉴定ALR-ELNs中下调[此处原文可能缺失某个基因或蛋白名称]表达的活性成分可以推动神经炎症性疾病治疗药物的开发。
