miRNA-328-3p regulates ZO-1 expression and inhibits PEDV proliferation via the PLC-β1-PKC pathway

PLoS One. 2025 Jan 3;20(1):e0316074. doi: 10.1371/journal.pone.0316074. eCollection 2025.

Abstract

Porcine epidemic diarrhea virus (PEDV) is a significant pathogen affecting swine, causing severe economic losses worldwide. This study explores the regulatory role of miRNA-328-3p to ZO-1 expression and its impact on PEDV proliferation via the PLC-β1-PKC pathway in IPEC-J2 cells. We found that miRNA-328-3p can target ZO-1, influencing its expression and subsequently affecting the integrity of tight junctions in the cells. Overexpression of PLC-β1, combined with miRNA-328-3p silencing, enhanced ZO-1 expression, while PLC-β1 knockdown combined with miRNA-328-3p overexpression inhibited ZO-1 expression. Furthermore, PLC-β1 overexpression increased both viral genome expression and PEDV titers, whereas its silencing had the opposite effect. Notably, our data indicated a negative correlation between PLC-β1 and PKC expression, and PKC silencing attenuated the upregulatory effect of PLC-β1 on ZO-1. These findings suggest that PLC-β1 modulates ZO-1 expression through the PKC pathway, providing new insights into the molecular mechanisms of PEDV infection and potential therapeutic targets.

MeSH terms

  • Animals
  • Cell Line
  • MicroRNAs* / genetics
  • MicroRNAs* / metabolism
  • Phospholipase C beta* / genetics
  • Phospholipase C beta* / metabolism
  • Porcine epidemic diarrhea virus* / genetics
  • Porcine epidemic diarrhea virus* / physiology
  • Protein Kinase C* / genetics
  • Protein Kinase C* / metabolism
  • Signal Transduction*
  • Swine
  • Virus Replication
  • Zonula Occludens-1 Protein* / genetics
  • Zonula Occludens-1 Protein* / metabolism

Substances

  • MicroRNAs
  • Zonula Occludens-1 Protein
  • Protein Kinase C
  • Phospholipase C beta

Grants and funding

The present work was supported by the National Natural Science Foundation of China (32072857, China); the Natural Science Foundation of Jilin Province, China (grant no. 20230101257JC); and the Natural Science Foundation of Jilin Province, China (grant no. 20240101219). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.