Citation

  • Authors: Woo SJ. et al.
  • Year: 2022
  • Journal: Int J Mol Sci 23 2999
  • Applications: in vivo / mimic miRNA / in vivo-jetPEI

Method

Before miRNA 148a mimic treatment, 106 CFUs BCG Pasteur in 100 µL saline into the intrapleural cavity was performed on day 0 to create a BCG-induced pleurisy animal model. To determine the effect of miR-148a on in vivo tuberculous fibrosis, a control mimic or miR-148a mimic (20 nmol/kg) combined with 100 µL of in vivo-JetPEI (PolyPlus Transfection, New York, NY, USA) transfection reagent was injected intravenously once every two days from day 0 in a total of 14 days. On day 15, all mice were sacrificed, and lung tissues and blood serum were collected.

Abstract

Extracellular matrix production by pleural mesothelial cells in response to Mycobacterium tuberculosis contributes to tuberculous fibrosis. NOX4 is involved in the pathogenesis of tuberculous fibrosis. In this study, we evaluated whether NOX4 gene-targeting microRNAs showed protective effects in tuberculosis fibrosis. TargetScan prediction software was used to identify candidate microRNAs that bind the 3' UTRs of NOX4, and microRNA-148a (miR-148a) was selected as the best miRNA candidate. A repressed and forced expression assay in Met5A cells was performed to investigate the causal relationship between miR-148a and NOX4. The role of miR-148a in tuberculous pleural fibrosis was studied using a murine model of Mycobacterium bovis bacillus Calmette-Guérin (BCG) pleural infection. Heat-killed M. tuberculosis (HKMT) induces NOX4 and POLDIP2 expression. We demonstrated the inhibitory effect of miR-148a on NOX4 and POLDIP2 expression. The increased expression of miR-148a suppressed HKMT-induced collagen-1A synthesis in PMC cells. In the BCG pleurisy model, miR-148a significantly reduced fibrogenesis and epithelial mesenchymal transition. High levels of miR-148a in tuberculous pleural effusion can be interpreted as a self-limiting homeostatic response. Our data indicate that miR-148a may protect against tuberculous pleural fibrosis by regulating NOX4 and POLDIP2.

Pubmed