Down-Regulation of miR-101 Contributes to Rheumatic Heart Disease Through Up-Regulating TLR2

Med Sci Monit. 2015 May 25:21:1500-6. doi: 10.12659/MSM.893540.

Abstract

Background: RHD is an autoimmune disease that arises following infection by S. pyogenes and imposes a heavy burden on public health.

Material and methods: We detected 11 selected miRNAs expressed in the cardiac tissues of 11 RHD patients and 11 controls. By employing dual-luciferase assay and Western blot, we identified the relationship between TLR2 and miR-101 and miR-101. We used ELISA to test the concentration of TNF-α, IL-1β, and IL-6.

Results: In cardiac tissue of RHD patients, miR-101 was significantly down-regulated (p=0.011). Ectopically expressed miR-101 repressed the luciferase activity by 27% through targeting TLR2 3'UTR. Combined with the results of Western blot, we confirmed that TLR2 is a direct target gene of miR-101. miR-101 knock-down is related to over-stimulated immune response in PGN-activated THP-1 cells. We detected a significantly higher concentration of TNF-α (p=0.0017), IL-1β (p=0.015), and IL-6 (p=0.014) in serum samples. TLR2 had a higher expression in patients in the protein level rather than the mRNA level, indicating that post-transcriptional regulation factors play a crucial role in regulating TLR2 expression.

Conclusions: The present study confirmed that miR-101 targets TLR2 3'UTR and represses TLR2 expression. This work also found an association between down-regulated miR-101 and rheumatic heart disease.

MeSH terms

  • Blotting, Western
  • Enzyme-Linked Immunosorbent Assay
  • Gene Expression Regulation / genetics
  • Gene Expression Regulation / physiology*
  • Humans
  • Interleukin-1beta / blood
  • Interleukin-6 / blood
  • Luciferases
  • MicroRNAs / metabolism*
  • Myocardium / metabolism*
  • Rheumatic Heart Disease / etiology*
  • Toll-Like Receptor 2 / metabolism*
  • Tumor Necrosis Factor-alpha / blood

Substances

  • Interleukin-1beta
  • Interleukin-6
  • MIRN101 microRNA, human
  • MicroRNAs
  • TLR2 protein, human
  • Toll-Like Receptor 2
  • Tumor Necrosis Factor-alpha
  • Luciferases