title: Down-regulation of miR-186 contributes to podocytes apoptosis in membranous nephropathy.
abstract: OBJECTIVE
  Podocytes apoptosis is the key process in the development of membranous nephropathy and miR-186 is reported to be related with cell apoptosis. Here we investigated the expression of miR-186 in membranous nephropathy (MGN) patients and the mechanism underlying the podocytes apoptosis.
  
  
  METHODS
  Thirty patients with MGN and 30 healthy people were included in this study. The expression of miR-186 was detected in renal tissue and podocyte cells exposed to AngII by real-time PCR. Caspase-3 activity was used to evaluate podocytes apoptosis. TLR4 and P2×7 protein expression was quantified by western blotting. miR-186 inhibitor and miR-186 mimic were transfected into cells to investigate the mechanism underlying miR-186 in podocytes apoptosis.
  
  
  RESULTS
  In MGN patients, the level of miR-186 was significantly down-regulated as well as the protein expression of TLR4 and P2×7 was up-regulated in renal tissue. In vitro experiments, TLR4 siRNA increased the expression of miR-186 and miR-186 inhibitor elevated the mRNA and protein expression of P2×7 in podocytes exposed to AngII. In addition, the level of cleaved-caspase-3 was up-regulated by miR-186 inhibitor. The TUNEL-positive cells and caspase-3 activity of podocytes induced by AngII were down-regulated by miR-186 mimic.
  
  
  CONCLUSIONS
  We revealed that TLR4 is involved in the regulation of miR-186 expression, and the anti-apoptotic effect of miR-186 on podocytes is correlated with P2×7 regulation.
authors.affiliation: Department of Nephrology, The First Affiliated Hospital of Soochow University, Suzhou 215006, PR China.
authors.forename: Wen-gang
authors.initials: WG
authors.lastname: Sha
authors.affiliation: Department of Nephrology, The First Affiliated Hospital of Soochow University, Suzhou 215006, PR China.
authors.forename: Lei
authors.initials: L
authors.lastname: Shen
authors.affiliation: Department of Nephrology, The First Affiliated Hospital of Soochow University, Suzhou 215006, PR China.
authors.forename: Ling
authors.initials: L
authors.lastname: Zhou
authors.affiliation: Department of Nephrology, The First Affiliated Hospital of Soochow University, Suzhou 215006, PR China.
authors.forename: De-yu
authors.initials: DY
authors.lastname: Xu
authors.affiliation: Department of Nephrology, The First Affiliated Hospital of Soochow University, Suzhou 215006, PR China. Electronic address: guoyuan_l@163.com.
authors.forename: Guo-yuan
authors.initials: GY
authors.lastname: Lu
mesh_terms: D020413:3' Untranslated Regions; D000804:Angiotensin II / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects; D001665:Binding Sites; D016022:Case-Control Studies; D053148:Caspase 3 / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D015536:Down-Regulation; D015433:Glomerulonephritis, Membranous / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D006801:Humans; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D050199:Podocytes / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D034622:RNA Interference; D058486:Receptors, Purinergic P2X7 / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D051197:Toll-Like Receptor 4 / Q000235:genetics / Q000378:metabolism; D014162:Transfection
chemical_list: D020413:3' Untranslated Regions; C533096:MIRN186 microRNA, human; D035683:MicroRNAs; C546468:P2RX7 protein, human; D058486:Receptors, Purinergic P2X7; C495345:TLR4 protein, human; D051197:Toll-Like Receptor 4; D000804:Angiotensin II; C505407:CASP3 protein, human; D053148:Caspase 3
keywords: Membranous nephropathy; P2×7; Podocyte; Toll like receptor; miR-186