Small RNAs in Exosome-like nanovesicles from Peucedanum japonicum (PjELNs)

Inflammation plays a key role in the pathogenesis of various diseases, including inflammatory bowel diseases such as ulcerative colitis and Crohn's disease. Similar to animals, plant cells produce exosome-like nanovesicles (ELNs) that play a role in transmitting biological signals from specific types of cells or tissues to other cells or tissues. Here we show that ELNs derived from Peucedanum japonicum (PjELNs) exhibit potent anti-inflammatory effects on macrophages and are effective in a mouse colitis model induced by dextran sodium sulfate (DSS). RNA sequence analysis reveals that PjELNs suppress multiple inflammatory cytokine-mediated signaling pathways in LPS-stimulated macrophage cell lines. We further show that PjENLs promote the differentiation of goblet cells, thereby enhancing mucin production and providing a protective effect on mucosal damage in a DSS-induced murine model of colitis. We also demonstrate that PjELNs inhibit the DSS-induced reduction in the number of transit amplifying cells. PjELNs exhibit anti-inflammatory properties, partly due to the presence of miRNA-like small RNAs that directly regulate the expression of the inflammatory cytokine IL6. Importantly, these small RNAs exhibit cross-species effects in both humans and mice. These findings reveal the mechanism by which plant-derived ELNs modulate inflammatory responses, suggesting their potential as a preventive and therapeutic strategy for inflammatory diseases of the colon. Overall design: Profiling of small RNAs, including miRNAs, in Exosome-like nanovesicles from Peucedanum japonicum (PjELNs)

炎症在包括溃疡性结肠炎、克罗恩病在内的多种炎症性肠病的发病机制中发挥关键作用。与动物细胞类似，植物细胞可产生外泌体样纳米囊泡（exosome-like nanovesicles，ELNs），这类囊泡可介导特定细胞或组织向其他细胞或组织传递生物信号。本研究发现，来自白花前胡（Peucedanum japonicum）的ELNs（下称PjELNs）对巨噬细胞具有强效抗炎活性，且在葡聚糖硫酸钠（dextran sodium sulfate，DSS）诱导的小鼠结肠炎模型中展现出显著治疗效果。RNA测序分析显示，PjELNs可抑制脂多糖（lipopolysaccharide，LPS）刺激的巨噬细胞系中多种炎症细胞因子介导的信号通路。进一步研究表明，PjELNs能够促进杯状细胞分化，进而增强黏蛋白产生，对DSS诱导的小鼠结肠炎模型中的黏膜损伤起到保护作用。本研究还证实，PjELNs可抑制DSS诱导的短暂扩增细胞（transit amplifying cells）数量减少。PjELNs的抗炎特性部分源于其携带的miRNA样小RNA，这类小RNA可直接调控炎症细胞因子白细胞介素6（interleukin 6，IL6）的表达。值得注意的是，这些小RNA在人类与小鼠中均展现出跨物种调控活性。上述研究结果揭示了植物来源ELNs调控炎症反应的潜在机制，提示其有望作为结肠炎症性疾病的预防与治疗候选策略。整体实验设计：对白花前胡来源的外泌体样纳米囊泡（PjELNs）中包含miRNA在内的小RNA开展表达谱分析。