MiRNA-328-3p Regulates ZO-1 Expression and Inhibits PEDV Proliferation Via the PLC-β1-PKC Pathway

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2025 Jan 3

PMID 39752351

Authors

Han Zhao

Jiayu Zhang

Dengliang Li

Zhanding Cui

Jiuyuan Liu

Di Bao

Yiming Wei

Xinyue Zhang

Zhimin Wu

Tianqi Zhang

Kai Wang

Shushuai Yi

Wei Lian

Guixue Hu

Affiliations

Soon will be listed here.

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.

References

Kim S, Lee J, Jung J, Kim I, Hyun B, Kim H . Genetic characterization of porcine epidemic diarrhea virus in Korea from 1998 to 2013. Arch Virol. 2015; 160(4):1055-64. PMC: 7086719. DOI: 10.1007/s00705-015-2353-y. View

Puerta-Guardo H, Glasner D, Espinosa D, Biering S, Patana M, Ratnasiri K . Flavivirus NS1 Triggers Tissue-Specific Vascular Endothelial Dysfunction Reflecting Disease Tropism. Cell Rep. 2019; 26(6):1598-1613.e8. PMC: 6934102. DOI: 10.1016/j.celrep.2019.01.036. View

Ikenouchi J, Umeda K, Tsukita S, Furuse M, Tsukita S . Requirement of ZO-1 for the formation of belt-like adherens junctions during epithelial cell polarization. J Cell Biol. 2007; 176(6):779-86. PMC: 2064052. DOI: 10.1083/jcb.200612080. View

Zhao S, Gao J, Zhu L, Yang Q . Transmissible gastroenteritis virus and porcine epidemic diarrhoea virus infection induces dramatic changes in the tight junctions and microfilaments of polarized IPEC-J2 cells. Virus Res. 2014; 192:34-45. PMC: 7114495. DOI: 10.1016/j.virusres.2014.08.014. View

Zhou Y, Ren Z, Zhang S, Wang H, Wu S, Bao W . Analysis of Intestinal Mucosa Integrity and Gene Functions upon Porcine Epidemic Diarrhea Virus Infection in Pigs. Animals (Basel). 2021; 11(3). PMC: 8000733. DOI: 10.3390/ani11030644. View

Yang Y, Follo M, Cocco L, Suh P . The physiological roles of primary phospholipase C. Adv Biol Regul. 2013; 53(3):232-41. DOI: 10.1016/j.jbior.2013.08.003. View

Kocherhans R, Bridgen A, Ackermann M, Tobler K . Completion of the porcine epidemic diarrhoea coronavirus (PEDV) genome sequence. Virus Genes. 2001; 23(2):137-44. PMC: 7089135. DOI: 10.1023/a:1011831902219. View

Liang J, Xie M, Hou L, Wang H, Luo J, Sun J . miRNAs derived from milk small extracellular vesicles inhibit porcine epidemic diarrhea virus infection. Antiviral Res. 2023; 212:105579. DOI: 10.1016/j.antiviral.2023.105579. View

Li J, Li Y, Liu P, Wang X, Ma Y, Zhong Q . Porcine Epidemic Diarrhea Virus Infection Disrupts the Nasal Endothelial Barrier To Favor Viral Dissemination. J Virol. 2022; 96(9):e0038022. PMC: 9093128. DOI: 10.1128/jvi.00380-22. View

10.

Balda M, Matter K . Tight junctions as regulators of tissue remodelling. Curr Opin Cell Biol. 2016; 42:94-101. DOI: 10.1016/j.ceb.2016.05.006. View

11.

Reglero-Real N, Colom B, Bodkin J, Nourshargh S . Endothelial Cell Junctional Adhesion Molecules: Role and Regulation of Expression in Inflammation. Arterioscler Thromb Vasc Biol. 2016; 36(10):2048-2057. PMC: 5035539. DOI: 10.1161/ATVBAHA.116.307610. View

12.

Lin C, Saif L, Marthaler D, Wang Q . Evolution, antigenicity and pathogenicity of global porcine epidemic diarrhea virus strains. Virus Res. 2016; 226:20-39. PMC: 7111424. DOI: 10.1016/j.virusres.2016.05.023. View

13.

Behm-Ansmant I, Rehwinkel J, Izaurralde E . MicroRNAs silence gene expression by repressing protein expression and/or by promoting mRNA decay. Cold Spring Harb Symp Quant Biol. 2007; 71:523-30. DOI: 10.1101/sqb.2006.71.013. View

14.

Hu Y, Xie X, Yang L, Wang A . A Comprehensive View on the Host Factors and Viral Proteins Associated With Porcine Epidemic Diarrhea Virus Infection. Front Microbiol. 2021; 12:762358. PMC: 8688245. DOI: 10.3389/fmicb.2021.762358. View

15.

Lin S, Han Y, Jenkin K, Lee S, Sasaki M, Klapproth J . Lysophosphatidic Acid Receptor 1 Is Important for Intestinal Epithelial Barrier Function and Susceptibility to Colitis. Am J Pathol. 2017; 188(2):353-366. PMC: 5785539. DOI: 10.1016/j.ajpath.2017.10.006. View

16.

Zhao H, Yang J, Wang Q, Cui Z, Li D, Niu J . Exosomal miRNA-328-3p targets ZO-3 and inhibits porcine epidemic diarrhea virus proliferation. Arch Virol. 2022; 167(3):901-910. DOI: 10.1007/s00705-022-05364-3. View

17.

Cui Z, Liu J, Xie C, Wang T, Sun P, Wang J . High-throughput screening unveils nitazoxanide as a potent PRRSV inhibitor by targeting NMRAL1. Nat Commun. 2024; 15(1):4813. PMC: 11156899. DOI: 10.1038/s41467-024-48807-y. View

18.

Haas A, Zihni C, Krug S, Maraspini R, Otani T, Furuse M . ZO-1 Guides Tight Junction Assembly and Epithelial Morphogenesis via Cytoskeletal Tension-Dependent and -Independent Functions. Cells. 2022; 11(23). PMC: 9740252. DOI: 10.3390/cells11233775. View

19.

Curry S, Schwartz K, Yoon K, Gabler N, Burrough E . Effects of porcine epidemic diarrhea virus infection on nursery pig intestinal function and barrier integrity. Vet Microbiol. 2017; 211:58-66. DOI: 10.1016/j.vetmic.2017.09.021. View

20.

Luo X, Guo L, Zhang J, Xu Y, Gu W, Feng L . Tight Junction Protein Occludin Is a Porcine Epidemic Diarrhea Virus Entry Factor. J Virol. 2017; 91(10). PMC: 5411586. DOI: 10.1128/JVI.00202-17. View