title: Development of a stretch-induced neurotrauma model for medium-throughput screening in vitro: identification of rifampicin as a neuroprotectant.
abstract: We hypothesized that an in vitro, stretch-based model of neural injury may be useful to identify compounds that decrease the cellular damage in neurotrauma.
  
  
  
  We screened three neural cell lines (B35, RN33B and SH-SY5Y) subjected to two differentiation methods and selected all-trans-retinoic acid-differentiated B35 rat neuroblastoma cells subjected to rapid stretch injury, coupled with a subthreshold concentration of H2O2, for the screen. The model induced marked alterations in gene expression and proteomic signature of the cells and culminated in delayed cell death (LDH release) and mitochondrial dysfunction [reduced 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) conversion]. Follow-up studies utilized human stem cell-derived neurons subjected to rapid stretch injury.
  
  
  
  From screening of a composite library of 3500 drugs, five drugs (when applied in a post-treatment regimen relative to stretch injury) improved both LDH and MTT responses. The effects of rifampicin were investigated in further detail. Rifampicin reduced cell necrosis and apoptosis and improved cellular bioenergetics. In a second model (stretch injury in human stem cell-derived neurons), rifampicin pretreatment attenuated LDH release, protected against the loss of neurite length and maintained neuron-specific class III β-tubulin immunoreactivity.
  
  
  
  We conclude that the current model is suitable for medium-throughput screening to identify compounds with neuroprotective potential. Rifampicin, when applied either in pre- or post-treatment, improves the viability of neurons subjected to stretch injury and protects against neurite loss. Rifampicin may be a candidate for repurposing for the therapy of traumatic brain injury.
  
  
  
  This article is part of a themed section on Inventing New Therapies Without Reinventing the Wheel: The Power of Drug Repurposing. To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v175.2/issuetoc.
authors.affiliation: Department of Anesthesiology, University of Texas Medical Branch, Galveston, TX, USA.
authors.forename: Isabel
authors.initials: I
authors.lastname: López-García
authors.affiliation: Department of Anesthesiology, University of Texas Medical Branch, Galveston, TX, USA.
authors.forename: Domokos
authors.initials: D
authors.lastname: Gerő
authors.affiliation: Department of Anesthesiology, University of Texas Medical Branch, Galveston, TX, USA.
authors.forename: Bartosz
authors.initials: B
authors.lastname: Szczesny
authors.affiliation: Department of Anesthesiology, University of Texas Medical Branch, Galveston, TX, USA.
authors.forename: Petra
authors.initials: P
authors.lastname: Szoleczky
authors.affiliation: Department of Anesthesiology, University of Texas Medical Branch, Galveston, TX, USA.
authors.forename: Gabor
authors.initials: G
authors.lastname: Olah
authors.affiliation: Department of Anesthesiology, University of Texas Medical Branch, Galveston, TX, USA.
authors.forename: Katalin
authors.initials: K
authors.lastname: Módis
authors.affiliation: Department of Pharmacology, University of Texas Medical Branch, Galveston, TX, USA.
authors.forename: Kangling
authors.initials: K
authors.lastname: Zhang
authors.affiliation: Department of Neuroscience & Cell Biology, University of Texas Medical Branch, Galveston, TX, USA.
authors.forename: Jungling
authors.initials: J
authors.lastname: Gao
authors.affiliation: Department of Neuroscience & Cell Biology, University of Texas Medical Branch, Galveston, TX, USA.
authors.forename: Ping
authors.initials: P
authors.lastname: Wu
authors.affiliation: Department of Pharmacology, University of Texas Medical Branch, Galveston, TX, USA.
authors.forename: Lawrence C
authors.initials: LC
authors.lastname: Sowers
authors.affiliation: Department of Anesthesiology, University of Texas Medical Branch, Galveston, TX, USA.
authors.forename: Doug
authors.initials: D
authors.lastname: DeWitt
authors.affiliation: Department of Anesthesiology, University of Texas Medical Branch, Galveston, TX, USA.
authors.forename: Donald S
authors.initials: DS
authors.lastname: Prough
authors.affiliation: Department of Anesthesiology, University of Texas Medical Branch, Galveston, TX, USA.
authors.forename: Csaba
authors.initials: C
authors.lastname: Szabo
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D000070642:Brain Injuries, Traumatic / Q000188:drug therapy* / Q000378:metabolism; D016923:Cell Death / Q000187:drug effects; D045744:Cell Line, Tumor; D004195:Disease Models, Animal; D004353:Drug Evaluation, Preclinical / Q000379:methods; D006801:Humans; D006861:Hydrogen Peroxide; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008928:Mitochondria / Q000378:metabolism; D018696:Neuroprotective Agents / Q000494:pharmacology / Q000627:therapeutic use; D012293:Rifampin / Q000494:pharmacology* / Q000627:therapeutic use*; D013314:Stress, Mechanical; D013778:Tetrazolium Salts / Q000378:metabolism
chemical_list: C000598529:3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide; D018696:Neuroprotective Agents; D013778:Tetrazolium Salts; D006861:Hydrogen Peroxide; D007770:L-Lactate Dehydrogenase; D012293:Rifampin