Wang Wenjia, Jin Ximing, Shao Qingqing, Liu Tong, Liu Tianli, Zhao Xinwei, Xu Lijun, Gao Wen, Hu Liu, Chen Zhuo
Institute of Integrated Traditional Chinese and Western Medicine, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China.
Health Management Center, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China.
Heliyon. 2024 Mar 5;10(5):e27019. doi: 10.1016/j.heliyon.2024.e27019. eCollection 2024 Mar 15.
Genital herpes, primarily caused by HSV-2 infection, remains a widespread sexually transmitted ailment. Extracellular vesicles play a pivotal role in host-virus confrontation. Recent research underscores the influence of Chinese herbal prescriptions on extracellular vesicle production and composition. This study aims to probe the impact of JieZe-1 (JZ-1) on extracellular vesicle components, elucidating its mechanisms against HSV-2 infection via extracellular vesicles.
The JZ-1's anti-HSV-2 effects were assessed using CCK-8 assay. Extracellular vesicles were precisely isolated utilizing ultracentrifugation and subsequently characterized through TEM, NTA, and Western Blot analyses. The anti-HSV-2 activity of extracellular vesicles was gauged using CCK-8, Western Blot, and immunofluorescence. Additionally, high-throughput sequencing was employed to detect miRNAs from extracellular vesicles, unraveling the potential antiviral mechanisms of JZ-1.
Antiviral efficacy of JZ-1 was shown in VK2/E6E7, HeLa, and Vero cells. The samples extracted from cell supernatant by ultracentrifugation were identified as extracellular vesicles. In VK2/E6E7 cells, extracellular vesicles from JZ-1 group enhanced cell survival rates and diminished the expression of intracellular viral protein gD, contrasting with the inert effect of control group vesicles. Extracellular vesicles from JZ-1 treated Vero cells demonstrated a weaker yet discernible anti-HSV-2 effect. Conversely, extracellular vesicles of HeLa cells exhibited no anti-HSV-2 effect from either group. High-throughput sequencing of VK2/E6E7 cell extracellular vesicles unveiled significant upregulation of miRNA-101, miRNA-29a, miRNA-29b, miRNA-29c, and miRNA-637 in JZ-1 group vesicles. KEGG pathway analysis suggested that these miRNAs may inhibit PI3K/AKT/mTOR signaling pathway and induce autophagy of host cells to protect against HSV-2. Western blot confirmed the induction of autophagy and inhibition of AKT/mTOR in VK2/E6E7 cells with JZ-1 group extracellular vesicles treatment.
JZ-1 had an anti-HSV-2 efficacy. After JZ-1 stimulation, VK2/E6E7 cells secreted extracellular vesicles which protect host cells from HSV-2 infection. High-throughput sequencing showed that these extracellular vesicles contained a large number of miRNAs targeting PI3K/AKT/mTOR pathway. JZ-1 group extracellular vesicles could inhibit the activation of AKT/mTOR pathway and induce the host cells autophagy.
生殖器疱疹主要由单纯疱疹病毒2型（HSV - 2）感染引起，仍是一种广泛传播的性传播疾病。细胞外囊泡在宿主与病毒的对抗中起关键作用。近期研究强调了中药方剂对细胞外囊泡产生和组成的影响。本研究旨在探究洁泽 - 1（JZ - 1）对细胞外囊泡成分的影响，阐明其通过细胞外囊泡对抗HSV - 2感染的机制。
使用CCK - 8法评估JZ - 1的抗HSV - 2作用。利用超速离心精确分离细胞外囊泡，随后通过透射电子显微镜（TEM）、纳米颗粒跟踪分析（NTA）和蛋白质免疫印迹法（Western Blot）进行表征。使用CCK - 8、Western Blot和免疫荧光法测定细胞外囊泡的抗HSV - 2活性。此外，采用高通量测序检测细胞外囊泡中的微小RNA（miRNA），以揭示JZ - 1潜在的抗病毒机制。
JZ - 1在VK2/E6E7、HeLa和Vero细胞中显示出抗病毒效果。通过超速离心从细胞上清液中提取的样品被鉴定为细胞外囊泡。在VK2/E6E7细胞中，JZ - 1组的细胞外囊泡提高了细胞存活率并降低了细胞内病毒蛋白gD的表达，与对照组囊泡的惰性作用形成对比。JZ - 1处理的Vero细胞的细胞外囊泡表现出较弱但可辨别的抗HSV - 2作用。相反，HeLa细胞的细胞外囊泡在两组中均未表现出抗HSV - 2作用。对VK2/E6E7细胞外囊泡进行高通量测序发现，JZ - 1组囊泡中miRNA - 101、miRNA - 29a、miRNA - 29b、miRNA - 29c和miRNA - 637显著上调。京都基因与基因组百科全书（KEGG）通路分析表明，这些miRNA可能抑制PI3K/AKT/mTOR信号通路并诱导宿主细胞自噬以抵御HSV - 2。蛋白质免疫印迹法证实，用JZ - 1组细胞外囊泡处理的VK2/E6E7细胞中自噬被诱导且AKT/mTOR被抑制。
JZ - 1具有抗HSV - 2效果。JZ - 1刺激后，VK2/E6E7细胞分泌细胞外囊泡，保护宿主细胞免受HSV - 2感染。高通量测序表明，这些细胞外囊泡含有大量靶向PI3K/AKT/mTOR通路的miRNA。JZ -
