title: TGF-β2 stimulates Tenon's capsule fibroblast proliferation in patients with glaucoma via suppression of miR-29b expression regulated by Nrf2.
abstract: OBJECTIVE
  To investigate the role of transforming growth factor-β2 (TGF-β2) in Tenon's capsule fibroblasts proliferation from glaucoma patients and the effect of nuclear factor (erythroid-derived 2)-like 2 (Nrf2) and miR-29b mRNA in this process.
  
  
  METHODS
  Tenon's capsule fibroblasts obtained from patients who had undergone selective glaucoma surgery (GTFs) were cultured and stimulated with 5 ng/mL TGF-β2 for 1, 3, 5, and 7 days. MTS assay was performed to detect the cell viability. Expression of Nrf2 and miR-29b was analyzed with western blot, RT-PCR and Chromatin immunoprecipitation assay (ChIP) in human fibroblast SX1412-B exposed to TGF-β2.
  
  
  RESULTS
  MTS assay showed that TGF-β2 was more stimulatory on GTFs proliferation than controls. At the same time, TGF-β2 exerted an intenser effect of decreasing the Nrf2 protein and miR-29b mRNA levels in GTFs, and the level of miR-29b was effectively regulated by Ad-Nrf2. In addition, ChIP assay suggested that TGF-β2 down-regulated miR-29b expression through repressing the binding of Nrf2 to the promoter of miR-29b. Finally, we found that overexpression Nrf2 in GTFs reduced the proliferation effect on GTFs induced by TGF-β2, while miR-29b inhibitor reversed this effect.
  
  
  CONCLUSIONS
  This study suggests that TGF-β2 has a time-effect relationship with Tenon's capsule fibroblasts proliferation from glaucoma patients, and it stimulates Tenon's capsule fibroblast proliferation via suppression of miR-29b expression regulated by Nrf2.
authors.affiliation: Department of Ophthalmology, Zhengzhou Central Hospital Zhengzhou 450007, China.
authors.forename: Wenying
authors.initials: W
authors.lastname: Ran
authors.affiliation: Department of Ophthalmology, Zhengzhou Central Hospital Zhengzhou 450007, China.
authors.forename: Dongmei
authors.initials: D
authors.lastname: Zhu
authors.affiliation: Department of Ophthalmology, Zhengzhou Central Hospital Zhengzhou 450007, China.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Feng
mesh_terms: D000328:Adult; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects* / Q000235:genetics; D002478:Cells, Cultured; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D005786:Gene Expression Regulation; D005901:Glaucoma / Q000378:metabolism*; D006801:Humans; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D008875:Middle Aged; D051267:NF-E2-Related Factor 2 / Q000235:genetics / Q000378:metabolism*; D058475:Tenon Capsule / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D053781:Transforming Growth Factor beta2 / Q000494:pharmacology*; D055815:Young Adult
chemical_list: C532045:MIRN29a microRNA, human; D035683:MicroRNAs; D051267:NF-E2-Related Factor 2; C495635:NFE2L2 protein, human; D053781:Transforming Growth Factor beta2
keywords: Nrf2; Tenon’s capsule fibroblast; Transforming growth factor-β2; glaucoma; miR-29b; proliferation