Bee venom promotes exosome secretion and alters miRNA cargo in T cells

In this study, we investigated the effects of bee venom (BV), a natural compound with known immunomodulatory activity, on the exosomal miRNA profile in human Jurkat T cells. The non-cytotoxic concentration of BV (2 μg/mL) was identified using the CCK-8 assay. Exosomes were isolated from BV-treated and control cells and characterized by transmission electron microscopy (TEM), Western blotting, and nanoparticle tracking analysis (NTA), with NTA also used to quantify particle concentration for assessing changes in secretion levels. High-throughput sequencing identified 74 differentially expressed miRNAs (44 upregulated, 30 downregulated), which were validated by qRT-PCR. Functional enrichment analyses (GO, KEGG, DO, Reactome) revealed associations with pathways related to neural development, cell cycle regulation, and tumorigenesis. BV treatment significantly promoted exosome release and selectively altered miRNA cargo.本研究探究了蜂毒（bee venom, BV）——一种已被证实具有免疫调节活性的天然化合物——对人Jurkat T细胞中外泌体miRNA表达谱的影响。本研究通过CCK-8试剂盒法（CCK-8 assay）确定了BV的无细胞毒性浓度（2 μg/mL）。研究人员从经BV处理的细胞及对照组细胞中分离出外泌体，并通过透射电子显微镜（transmission electron microscopy, TEM）、蛋白质免疫印迹（Western blotting）以及纳米颗粒追踪分析（nanoparticle tracking analysis, NTA）对外泌体进行表征；同时利用NTA对颗粒浓度进行定量，以评估外泌体分泌水平的变化。高通量测序（High-throughput sequencing）共筛选出74个差异表达miRNA，其中44个表达上调、30个表达下调，随后通过实时定量逆转录聚合酶链反应（qRT-PCR）对上述筛选结果进行了验证。功能富集分析涵盖基因本体（Gene Ontology, GO）、京都基因与基因组百科全书（Kyoto Encyclopedia of Genes and Genomes, KEGG）、疾病本体（Disease Ontology, DO）及Reactome数据库，分析结果显示，差异表达miRNA所富集的通路与神经发育、细胞周期调控及肿瘤发生密切相关。BV处理可显著促进外泌体的分泌，并选择性地改变外泌体所携带的miRNA载荷。