title: NBHA reduces acrolein-induced changes in ARPE-19 cells: possible involvement of TGFβ.
abstract: OBJECTIVE
  Acrolein, a toxic, reactive aldehyde formed metabolically and environmentally, has been implicated in the damage to and dysfunction of the retinal pigment epithelium (RPE) that accompanies age-related macular degeneration (AMD). Our purpose was to investigate the potential of acrolein to influence the release of transforming growth factor beta-2 (TGFβ2) and vascular endothelial growth factor (VEGF), to assess the ability of N-benzylhydroxylamine (NBHA) to prevent the effect of acrolein on cytokine release and reduction of viable cells, and to explore the pathway by which acrolein might be causing the increase of VEGF.
  
  
  METHODS
  Confluent ARPE-19 cells were treated with acrolein and/or NBHA. They were also pretreated with SIS3, a specific inhibitor of SMAD 3, and ZM39923, a JAK3 inhibitor, before being treated with acrolein. Viable cells were counted; ELISA was used to measure the TGFβ2 and/or VEGF in the conditioned media.
  
  
  RESULTS
  Acrolein was shown to reduce the number of viable ARPE-19 cells and to upregulate the release of the proangiogenic cytokines TGFβ2 and VEGF. Co-treatment with 200 μM NBHA significantly reduced the effects of acrolein on viable cell number and TGFβ2 release. Pretreatment of the cells with SIS3 partially blocked the action of acrolein on decreased viable cell number and VEGF upregulation, suggesting that part of the effects of acrolein are mediated by the increased levels of TGFβ and its signaling.
  
  
  CONCLUSIONS
  Our results suggest that the action of acrolein on the reduction of viability and VEGF increase by ARPE-19 cells is partially mediated by TGFβ2. By reducing the effects of acrolein, NBHA and SIS3 could be potential pharmacological agents in the prevention and progression of acrolein-induced damage to the RPE that relates to AMD.
authors.affiliation: The University of Texas at San Antonio, San Antonio, Texas 78249, USA.
authors.forename: Eileen
authors.initials: E
authors.lastname: Vidro-Kotchan
authors.forename: Bharat Bhushan
authors.initials: BB
authors.lastname: Yendluri
authors.forename: Terrie
authors.initials: T
authors.lastname: Le-Thai
authors.forename: Andrew
authors.initials: A
authors.lastname: Tsin
mesh_terms: D000171:Acrolein / Q000037:antagonists & inhibitors* / Q000633:toxicity; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D006898:Hydroxylamines / Q000494:pharmacology*; D007546:Isoquinolines / Q000494:pharmacology; D011725:Pyridines / Q000494:pharmacology; D011758:Pyrroles / Q000494:pharmacology; D055213:Retinal Pigment Epithelium / Q000187:drug effects* / Q000378:metabolism; D053781:Transforming Growth Factor beta2 / Q000378:metabolism*; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism
chemical_list: C508504:6,7-dimethyl-2-(2E)-3-(1-methyl-2-phenyl-1H-pyrrolo(2,3-b)pyridin-3-yl-prop-2-enoyl)-1,2,3,4-tetrahydroisoquinoline hydrochloride; D006898:Hydroxylamines; D007546:Isoquinolines; D011725:Pyridines; D011758:Pyrroles; D053781:Transforming Growth Factor beta2; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; C037705:4-nitrobenzyloxyamine; D000171:Acrolein