Zhongguo shuxue zazhi (May 2022)

Effects of HBV-induced exosomes on macrophage phenotype and function

  • Rongrong MU,
  • Shuang LI,
  • Haiyan YE,
  • Limin CHEN,
  • Yujia LI,
  • Shilin LI

DOI
https://doi.org/10.13303/j.cjbt.issn.1004-549x.2022.05.004
Journal volume & issue
Vol. 35, no. 5
pp. 488 – 493

Abstract

Read online

Objective To investigate the effect of exosomes produced by hepatitis B virus (HBV)-infected cells on the phenotype and function of macrophages. Methods The exosomes secreted by HepAD38 cells, which were capable of producing HBV and HepG2 cells, were collected by ultracentrifugation combined with immunosorbent method.The quality and purity of the extracted exosomes were verified by nanoparticle tracking analysis (NTA), scanning electron microscope and Western blot.The M0 THP-1 macrophages differentiated by PMA were stimulated by HepAD38 derived- or HepG2 derived exosomes.Total RNA and protein samples were collected at different time points after stimulation.Real-time fluorescence quantitative PCR (qRT-PCR) and enzyme-linked immunosorbent assay (ELISA) were used to detect cytokine mRNA and protein expressions, respectively.Meanwhile, neutral red assay was performed to analyze macrophage pinocytosis activity, and a commercial kit was used to measure reactive oxygen species (ROS) in THP-1 macrophages.Human reverse transcription chip detection was performed to obtain the microRNAs profile of the exosomes.And the effect of selected miRNA on macrophages was further confirmed by qRT-PCR. Results Compared with HepG2-derived exosomes, HepAD38-derived exosomes increased the mRNA and protein expressions of IL-1β, MCP-1 and TNFα significantly.However, no difference of pinocytosis capacity or ROS production was found between the HepAD38-derived exosomes group and HepG2-derived exosomes group.Human reverse transcription chip detection results were verified by KEGG analysis and qRT-PCR, and it was found that miR-6824-3p could also significantly increase the expression levels of IL-1β, MCP-1 and TNFα after high expression. Conclusion This study found that exosomes produced by HepAD38 cells may stimulate macrophages to produce inflammatory factors such as IL-1β, MCP-1 and TNFα through miR-6824-3p, thereby playing a role in HBV infection.

Keywords