Department of Pharmacy, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei Province, People's Republic of China.
The Center for Biomedical Research, NHC Key Laboratory of Respiratory Diseases, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei Province, People's Republic of China.
Int J Nanomedicine. 2023 Oct 24;18:5983-6000. doi: 10.2147/IJN.S425173. eCollection 2023.
Pathogenic respiratory RNA viruses, including influenza A virus (IAV), respiratory syncytial virus (RSV), and SARS-CoV-2, are major causes of causes of acute respiratory infection globally. Plant-derived exosome-like nanoparticles containing miRNAs have shown substantial cross-kingdom regulatory effects on both viral and human transcripts. (), a traditional Chinese medicine frequently used to treat respiratory diseases. However, the role of derived exosome-like nanoparticles (HELNs) and the miRNA they encapsulated are unclear.
HELNs were isolated from fresh underground roots (uHELNs) and above ground stems and leaves (aHELNs) using differential centrifugation. The HELNs were identified using transmission electron microscopy, nanoparticle tracking analysis, and zeta potential. Small RNA sequencing and RT-PCR were employed to determine the miRNA expression in uHELNs and aHELNs. All genomes were sourced from the NCBI database. Target prediction of viral genomes was performed using RNAhybrid, while human target prediction was conducted using both RNAhybrid and Miranda. Functional enrichment analysis was applied to the predicted human targets to explore the hub targets and their roles in antiviral effects. The accessibility of miRNA target sites was determined through the MFOLD web server, and customized dual-luciferase reporter assays were administered to validate the computational findings.
A total of 12 highly enriched miRNAs were identified in both uHELNs and aHELNs. Upon prediction and verification, miR858a and miR858b were shown to target the gene in H1N1, while miR166a-3p targeted the in SARS-CoV-2. However, no valid miRNA targets were found for RSV. Regarding human transcripts, miR168a-3p, miR168b-3p, and miR8175 were found to inhibit expression, and novel_mir2 could suppress both and expression.
This study sheds light on the collaborative antiviral mechanism of miRNAs in HELNs across two species and explores the potential antiviral scopes of both miRNAs and HELNs.
致病性呼吸道 RNA 病毒，包括甲型流感病毒（IAV）、呼吸道合胞病毒（RSV）和 SARS-CoV-2，是全球急性呼吸道感染的主要原因。含有 miRNA 的植物衍生外泌体样纳米颗粒对病毒和人类转录物都表现出显著的跨物种调控作用。（）是一种常用于治疗呼吸道疾病的中药。然而，其衍生的外泌体样纳米颗粒（HELNs）及其包裹的 miRNA 的作用尚不清楚。
使用差速离心法从新鲜地下根（uHELNs）和地上茎和叶（aHELNs）中分离 HELNs。通过透射电子显微镜、纳米颗粒跟踪分析和zeta 电位对 HELNs 进行鉴定。使用小 RNA 测序和 RT-PCR 确定 uHELNs 和 aHELNs 中的 miRNA 表达。所有基因组均来自 NCBI 数据库。使用 RNAhybrid 对病毒基因组进行靶预测，同时使用 RNAhybrid 和 Miranda 对人类靶标进行预测。对预测的人类靶标进行功能富集分析，以探索抗病毒作用中的关键靶标及其作用。通过 MFOLD 网络服务器确定 miRNA 靶位的可及性，并进行定制的双荧光素酶报告基因检测以验证计算结果。
在 uHELNs 和 aHELNs 中均鉴定出 12 种高度富集的 miRNA。通过预测和验证，miR858a 和 miR858b 被证明靶向 H1N1 的 基因，而 miR166a-3p 靶向 SARS-CoV-2 的 基因。然而，对于 RSV，没有发现有效的 miRNA 靶标。对于人类转录物，miR168a-3p、miR168b-3p 和 miR8175 被发现抑制 表达，而 novel_mir2 可以抑制 和 表达。
本研究揭示了 HELNs 中 miRNA 在两种物种之间的协同抗病毒机制，并探讨了 miRNAs 和 HELNs 的潜在抗病毒范围。
