沿肠道-肝门静脉-肝轴的 LPS+ 细菌胞外囊泡的特征。

IF 15.5 1区 医学 Q1 CELL BIOLOGY Journal of Extracellular Vesicles Pub Date : 2024-07-13 DOI:10.1002/jev2.12474
Heetanshi Jain, Ashish Kumar, Sameh Almousa, Shalini Mishra, Kendall L. Langsten, Susy Kim, Mitu Sharma, Yixin Su, Sangeeta Singh, Bethany A. Kerr, Gagan Deep
{"title":"沿肠道-肝门静脉-肝轴的 LPS+ 细菌胞外囊泡的特征。","authors":"Heetanshi Jain,&nbsp;Ashish Kumar,&nbsp;Sameh Almousa,&nbsp;Shalini Mishra,&nbsp;Kendall L. Langsten,&nbsp;Susy Kim,&nbsp;Mitu Sharma,&nbsp;Yixin Su,&nbsp;Sangeeta Singh,&nbsp;Bethany A. Kerr,&nbsp;Gagan Deep","doi":"10.1002/jev2.12474","DOIUrl":null,"url":null,"abstract":"<p>Gut microbiome dysbiosis is a major contributing factor to several pathological conditions. However, the mechanistic understanding of the communication between gut microbiota and extra-intestinal organs remains largely elusive. Extracellular vesicles (EVs), secreted by almost every form of life, including bacteria, could play a critical role in this inter-kingdom crosstalk and are the focus of present study. Here, we present a novel approach for isolating lipopolysaccharide (LPS)+ bacterial extracellular vesicles (bEV<sup>LPS</sup>) from complex biological samples, including faeces, plasma and the liver from lean and diet-induced obese (DIO) mice. bEV<sup>LPS</sup> were extensively characterised using nanoparticle tracking analyses, immunogold labelling coupled with transmission electron microscopy, flow cytometry, super-resolution microscopy and 16S sequencing. In liver tissues, the protein expressions of TLR4 and a few macrophage-specific biomarkers were assessed by immunohistochemistry, and the gene expressions of inflammation-related cytokines and their receptors (<i>n</i> = 89 genes) were measured using a PCR array. Faecal samples from DIO mice revealed a remarkably lower concentration of total EVs but a significantly higher percentage of LPS+ EVs. Interestingly, DIO faecal bEV<sup>LPS</sup> showed a higher abundance of <i>Proteobacteria</i> by 16S sequencing. Importantly, in DIO mice, a higher number of total EVs and bEV<sup>LPS</sup> consistently entered the hepatic portal vein and subsequently reached the liver, associated with increased expression of TLR4, macrophage markers (F4/80, CD86 and CD206), cytokines and receptors (<i>Il1rn</i>, <i>Ccr1</i>, <i>Cxcl10</i>, <i>Il2rg</i> and <i>Ccr2</i>). Furthermore, a portion of bEV<sup>LPS</sup> escaped liver and entered the peripheral circulation. In conclusion, bEV could be the key mediator orchestrating various well-established biological effects induced by gut bacteria on distant organs.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":null,"pages":null},"PeriodicalIF":15.5000,"publicationDate":"2024-07-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jev2.12474","citationCount":"0","resultStr":"{\"title\":\"Characterisation of LPS+ bacterial extracellular vesicles along the gut-hepatic portal vein-liver axis\",\"authors\":\"Heetanshi Jain,&nbsp;Ashish Kumar,&nbsp;Sameh Almousa,&nbsp;Shalini Mishra,&nbsp;Kendall L. Langsten,&nbsp;Susy Kim,&nbsp;Mitu Sharma,&nbsp;Yixin Su,&nbsp;Sangeeta Singh,&nbsp;Bethany A. Kerr,&nbsp;Gagan Deep\",\"doi\":\"10.1002/jev2.12474\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p>Gut microbiome dysbiosis is a major contributing factor to several pathological conditions. However, the mechanistic understanding of the communication between gut microbiota and extra-intestinal organs remains largely elusive. Extracellular vesicles (EVs), secreted by almost every form of life, including bacteria, could play a critical role in this inter-kingdom crosstalk and are the focus of present study. Here, we present a novel approach for isolating lipopolysaccharide (LPS)+ bacterial extracellular vesicles (bEV<sup>LPS</sup>) from complex biological samples, including faeces, plasma and the liver from lean and diet-induced obese (DIO) mice. bEV<sup>LPS</sup> were extensively characterised using nanoparticle tracking analyses, immunogold labelling coupled with transmission electron microscopy, flow cytometry, super-resolution microscopy and 16S sequencing. In liver tissues, the protein expressions of TLR4 and a few macrophage-specific biomarkers were assessed by immunohistochemistry, and the gene expressions of inflammation-related cytokines and their receptors (<i>n</i> = 89 genes) were measured using a PCR array. Faecal samples from DIO mice revealed a remarkably lower concentration of total EVs but a significantly higher percentage of LPS+ EVs. Interestingly, DIO faecal bEV<sup>LPS</sup> showed a higher abundance of <i>Proteobacteria</i> by 16S sequencing. Importantly, in DIO mice, a higher number of total EVs and bEV<sup>LPS</sup> consistently entered the hepatic portal vein and subsequently reached the liver, associated with increased expression of TLR4, macrophage markers (F4/80, CD86 and CD206), cytokines and receptors (<i>Il1rn</i>, <i>Ccr1</i>, <i>Cxcl10</i>, <i>Il2rg</i> and <i>Ccr2</i>). Furthermore, a portion of bEV<sup>LPS</sup> escaped liver and entered the peripheral circulation. In conclusion, bEV could be the key mediator orchestrating various well-established biological effects induced by gut bacteria on distant organs.</p>\",\"PeriodicalId\":15811,\"journal\":{\"name\":\"Journal of Extracellular Vesicles\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":15.5000,\"publicationDate\":\"2024-07-13\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jev2.12474\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of Extracellular Vesicles\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://onlinelibrary.wiley.com/doi/10.1002/jev2.12474\",\"RegionNum\":1,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"CELL BIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Extracellular Vesicles","FirstCategoryId":"3","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/jev2.12474","RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"CELL BIOLOGY","Score":null,"Total":0}
引用次数: 0

摘要

肠道微生物群失调是导致多种病症的一个主要因素。然而,人们对肠道微生物群与肠道外器官之间交流的机理仍然知之甚少。包括细菌在内的几乎所有生命形式都会分泌胞外囊泡 (EV),它们可能在这种跨领域的串联中发挥关键作用,这也是本研究的重点。在这里,我们提出了一种从复杂生物样本(包括粪便、血浆以及瘦小鼠和饮食诱导肥胖(DIO)小鼠的肝脏)中分离脂多糖(LPS)+细菌胞外囊泡(bEVLPS)的新方法。我们使用纳米颗粒跟踪分析、免疫金标记结合透射电子显微镜、流式细胞术、超分辨率显微镜和 16S 测序对 bEVLPS 进行了广泛表征。在肝脏组织中,通过免疫组化评估了 TLR4 和一些巨噬细胞特异性生物标志物的蛋白表达,并使用 PCR 阵列测量了炎症相关细胞因子及其受体(n = 89 个基因)的基因表达。DIO 小鼠的粪便样本显示,总 EVs 的浓度明显降低,但 LPS+ EVs 的比例明显升高。有趣的是,通过 16S 测序,DIO 粪便中的 bEVLPS 显示了更高的变形杆菌丰度。重要的是,在 DIO 小鼠中,更多的总 EVs 和 bEVLPS 持续进入肝门静脉并随后到达肝脏,这与 TLR4、巨噬细胞标记物(F4/80、CD86 和 CD206)、细胞因子和受体(Il1rn、Ccr1、Cxcl10、Il2rg 和 Ccr2)的表达增加有关。此外,一部分 bEVLPS 从肝脏排出,进入外周循环。总之,bEV 可能是协调肠道细菌对远处器官诱导的各种公认生物效应的关键介质。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

摘要图片

查看原文
分享 分享
微信好友 朋友圈 QQ好友 复制链接
本刊更多论文
Characterisation of LPS+ bacterial extracellular vesicles along the gut-hepatic portal vein-liver axis

Gut microbiome dysbiosis is a major contributing factor to several pathological conditions. However, the mechanistic understanding of the communication between gut microbiota and extra-intestinal organs remains largely elusive. Extracellular vesicles (EVs), secreted by almost every form of life, including bacteria, could play a critical role in this inter-kingdom crosstalk and are the focus of present study. Here, we present a novel approach for isolating lipopolysaccharide (LPS)+ bacterial extracellular vesicles (bEVLPS) from complex biological samples, including faeces, plasma and the liver from lean and diet-induced obese (DIO) mice. bEVLPS were extensively characterised using nanoparticle tracking analyses, immunogold labelling coupled with transmission electron microscopy, flow cytometry, super-resolution microscopy and 16S sequencing. In liver tissues, the protein expressions of TLR4 and a few macrophage-specific biomarkers were assessed by immunohistochemistry, and the gene expressions of inflammation-related cytokines and their receptors (n = 89 genes) were measured using a PCR array. Faecal samples from DIO mice revealed a remarkably lower concentration of total EVs but a significantly higher percentage of LPS+ EVs. Interestingly, DIO faecal bEVLPS showed a higher abundance of Proteobacteria by 16S sequencing. Importantly, in DIO mice, a higher number of total EVs and bEVLPS consistently entered the hepatic portal vein and subsequently reached the liver, associated with increased expression of TLR4, macrophage markers (F4/80, CD86 and CD206), cytokines and receptors (Il1rn, Ccr1, Cxcl10, Il2rg and Ccr2). Furthermore, a portion of bEVLPS escaped liver and entered the peripheral circulation. In conclusion, bEV could be the key mediator orchestrating various well-established biological effects induced by gut bacteria on distant organs.

求助全文
通过发布文献求助,成功后即可免费获取论文全文。 去求助
来源期刊
Journal of Extracellular Vesicles
Journal of Extracellular Vesicles Biochemistry, Genetics and Molecular Biology-Cell Biology
CiteScore
27.30
自引率
4.40%
发文量
115
审稿时长
12 weeks
期刊介绍: The Journal of Extracellular Vesicles is an open access research publication that focuses on extracellular vesicles, including microvesicles, exosomes, ectosomes, and apoptotic bodies. It serves as the official journal of the International Society for Extracellular Vesicles and aims to facilitate the exchange of data, ideas, and information pertaining to the chemistry, biology, and applications of extracellular vesicles. The journal covers various aspects such as the cellular and molecular mechanisms of extracellular vesicles biogenesis, technological advancements in their isolation, quantification, and characterization, the role and function of extracellular vesicles in biology, stem cell-derived extracellular vesicles and their biology, as well as the application of extracellular vesicles for pharmacological, immunological, or genetic therapies. The Journal of Extracellular Vesicles is widely recognized and indexed by numerous services, including Biological Abstracts, BIOSIS Previews, Chemical Abstracts Service (CAS), Current Contents/Life Sciences, Directory of Open Access Journals (DOAJ), Journal Citation Reports/Science Edition, Google Scholar, ProQuest Natural Science Collection, ProQuest SciTech Collection, SciTech Premium Collection, PubMed Central/PubMed, Science Citation Index Expanded, ScienceOpen, and Scopus.
期刊最新文献
A switch from lysosomal degradation to secretory autophagy initiates osteogenic bone metastasis in prostate cancer Correction to MAP kinase kinase 1 (MEK1) within extracellular vesicles inhibits tumour growth by promoting anti-tumour immunity A novel multiplexed immunoassay for surface-exposed proteins in plasma extracellular vesicles Extracellular vesicles from human-induced pluripotent stem cell-derived neural stem cells alleviate proinflammatory cascades within disease-associated microglia in Alzheimer's disease Real-time monitoring of small extracellular vesicles (sEVs) by in vivo flow cytometry
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
现在去查看 取消
×
提示
确定
0
微信
客服QQ
Book学术公众号 扫码关注我们
反馈
×
意见反馈
请填写您的意见或建议
请填写您的手机或邮箱
已复制链接
已复制链接
快去分享给好友吧!
我知道了
×
扫码分享
扫码分享
Book学术官方微信
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术
文献互助 智能选刊 最新文献 互助须知 联系我们:info@booksci.cn
Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。
Copyright © 2023 Book学术 All rights reserved.
ghs 京公网安备 11010802042870号 京ICP备2023020795号-1