Comparative study on characterization, miRNA composition, and target gene enrichment pathways of AstragaliRadix-derived extracellular vesicles-like particles obtained by soaking and water decoction pretreatment methods
YANG Wanpeng, XU Sinan, DONG Mengwei, LI Wenfang, BA Te, LIU Yang, FU Songtao
Affiliations
doi: 10.7501/j.issn.0253-2670.2026.14.015
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摘要
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目的 探究浸泡与水煎2种预处理方式所得黄芪来源囊泡样纳米颗粒（AstragaliRadix-derived extracellular vesicles-like particles，AR-EVLP）在表征、miRNA组成和靶基因富集通路上的差异。方法 通过透射电子显微镜、纳米颗粒跟踪分析仪、纳米激光粒度仪、蛋白浓度定量、银染、实时荧光定量PCR、miRNA测序及生物信息学分析，对比分析2种预处理所得AR-EVLP的形态、颗粒浓度、粒径大小、ζ电位、蛋白浓度、蛋白分布等表征差异及miRNA组成和分子功能差异。结果 浸泡预处理所得AR-EVLP质量优于水煎预处理；水煎预处理所得AR-EVLP的产率较高、粒径相对较大、ζ电位绝对值相对较小、蛋白浓度更高且分布更为集中。测序结果显示，2种预处理方式所得AR-EVLP有219个差异性表达miRNA。基因本体论（gene ontology，GO）分析显示，2种预处理所得AR-EVLP的miRNA靶基因均在蛋白结合条目显著富集，提示其跨物种调控的潜能。京都基因与基因组百科全书（Kyoto encyclopedia of genes and genomes，KEGG）富集分析显示，浸泡预处理所得AR-EVLP的miRNA中lja-miR1511-3p_R-3_1ss2AC和gma-miR1511_R+1_1ss2AC，可能通过甘油磷脂代谢通路在脂肪肝、神经系统退行性疾病具有治疗潜能；水煎预处理所得AR-EVLP的miRNA中mtr-miR164a、stu-miR477a-5p_R+1_1 ss21CG、gma-MIR162c-p5可能通过胶质瘤、前列腺癌、p53信号通路、Hedgehog信号通路、ErbB信号通路、丝裂原活化蛋白激酶（mitogen-activated protein kinase，MAPK）信号通路等通路治疗癌症相关疾病。结论 不同预处理所得AR-EVLP在表征、蛋白成分、miRNA表达谱及其生物学功能导向存在显著差异。浸泡预处理所得AR-EVLP质量较好，倾向于脂肪肝、神经系统退行性疾病等疾病治疗，水煎预处理所得AR-EVLP的产率较高，倾向于癌症相关疾病治疗。
关键词
黄芪
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囊泡样纳米颗粒
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提取技术
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miRNA
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生物信息学分析
Abstract
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Objective To explore the differences in characterization, miRNA composition, and target gene enrichment pathways of Astragali Radix-derived extracellular vesicle-like particles (AR-EVLP) obtained through two pretreatment methods: soaking and decoction. Methods Transmission electron microscopy, nanoparticle tracking analysis, nanoscale laser particle sizing, protein quantification, silver staining, real-time quantitative PCR, miRNA sequencing, and bioinformatics analysis were used to compare the morphology, particle concentration, size, ζ potential, protein concentration, protein distribution, miRNA composition, and molecular function differences of AR-EVLP obtained from the two pretreatment methods. Results The quality of AR-EVLP obtained from soaking was better than that obtained from decoction; AR-EVLP from decoction had a higher yield, relatively larger particle size, smaller absolute ζ potential, higher protein concentration, and more concentrated protein distribution. Sequencing results showed 219 differentially expressed miRNAs between the AR-EVLP obtained by the two pretreatment methods. Gene Ontology (GO) analysis indicated that the miRNA target genes from both pretreatment methods were significantly enriched in protein binding, suggesting potential cross-species regulatory effects. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis showed that miRNAs lja-miR1511-3p_R-3_1ss2AC and gma-miR1511_R+1_1ss2AC from AR-EVLP obtained by soaking might have therapeutic potential for fatty liver and neurodegenerative diseases via the glycerophospholipid metabolism pathway. miRNAs mtr-miR164a, stu-miR477a-5p_R+1_1ss21CG, and gma-MIR162c-p5 from AR-EVLP obtained by decoction might treat cancer-related diseases through pathways such as glioma, prostate cancer, p53 signaling, Hedgehog signaling, ErbB signaling, and mitogen-activated protein kinase (MAPK) signaling. Conclusion AR-EVLP obtained by different pretreatment methods exhibited significant differences in characterization, protein composition, miRNA expression profiles, and associated biological functions. AR-EVLP from soaking had better quality and tended to be useful for treating diseases like fatty liver and neurodegenerative disorders, while AR-EVLP from decoction had a higher yield and tended to be geared toward treating cancer-related diseases.
YANG Wanpeng, XU Sinan, DONG Mengwei, LI Wenfang, BA Te, LIU Yang, FU Songtao.
Comparative study on characterization, miRNA composition, and target gene enrichment pathways of AstragaliRadix-derived extracellular vesicles-like particles obtained by soaking and water decoction pretreatment methods[J].
Chinese Traditional and Herbal Drugs,
2026
, 57
(14)
: 5523
-5536
.
DOI: 10.7501/j.issn.0253-2670.2026.14.015
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