title: Down-Regulation of miR-101 Contributes to Rheumatic Heart Disease Through Up-Regulating TLR2.
abstract: BACKGROUND
  RHD is an autoimmune disease that arises following infection by S. pyogenes and imposes a heavy burden on public health.
  
  
  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.
authors.affiliation: Department of Critical Care Medicine, Affiliated Hospital of Qingdao University, Qingdao, Shandong, China (mainland).
authors.forename: Hai
authors.initials: H
authors.lastname: Dong
authors.affiliation: Department of Ultrasound, Affiliated Hospital of Qingdao University, Qingdao, Shandong, China (mainland).
authors.forename: Yongmei
authors.initials: Y
authors.lastname: Sun
authors.affiliation: Department of Critical Care Medicine, Affiliated Hospital of Qingdao University, Qingdao, Shandong, China (mainland).
authors.forename: Feng
authors.initials: F
authors.lastname: Shan
authors.affiliation: Department of Critical Care Medicine, Affiliated Hospital of Qingdao University, Qingdao, Shandong, China (mainland).
authors.forename: Qiang
authors.initials: Q
authors.lastname: Sun
authors.affiliation: Department of Critical Care Medicine, Affiliated Hospital of Qingdao University, Qingdao, Shandong, China (mainland).
authors.forename: Bingxin
authors.initials: B
authors.lastname: Yang
mesh_terms: D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D005786:Gene Expression Regulation / Q000235:genetics / Q000502:physiology*; D006801:Humans; D053583:Interleukin-1beta / Q000097:blood; D015850:Interleukin-6 / Q000097:blood; D008156:Luciferases; D035683:MicroRNAs / Q000378:metabolism*; D009206:Myocardium / Q000378:metabolism*; D012214:Rheumatic Heart Disease / Q000209:etiology*; D051195:Toll-Like Receptor 2 / Q000378:metabolism*; D014409:Tumor Necrosis Factor-alpha / Q000097:blood
chemical_list: D053583:Interleukin-1beta; D015850:Interleukin-6; C534072:MIRN101 microRNA, human; D035683:MicroRNAs; C495342:TLR2 protein, human; D051195:Toll-Like Receptor 2; D014409:Tumor Necrosis Factor-alpha; D008156:Luciferases