title: Bee venom effects on ubiquitin proteasome system in hSOD1(G85R)-expressing NSC34 motor neuron cells.
abstract: BACKGROUND
  Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease that results from a progressive loss of motor neurons. Familial ALS (fALS) is caused by missense mutations in Cu, Zn-superoxide dismutase 1 (SOD1) that frequently result in the accumulation of mutant protein aggregates that are associated with impairments in the ubiquitin-proteasome system (UPS). UPS impairment has been implicated in many neurological disorders. Bee venom (BV) extracted from honey bees has been used as a traditional medicine for treating inflammatory diseases and has been shown to attenuate the neuroinflammatory events that occur in a symptomatic ALS animal model.
  
  
  METHODS
  NSC34 cells were transiently transfected with a WT or G85R hSOD1-GFP construct for 24 hrs and then stimulated with 2.5 μg/ml BV for 24 hrs. To determine whether a SOD1 mutation affects UPS function in NSC34 cells, we examined proteasome activity and performed western blotting and immunofluorescence using specific antibodies, such as anti-misfolded SOD1, anti-ubiquitin, anti-GRP78, anti-LC3, and anti-ISG15 antibodies.
  
  
  RESULTS
  We found that GFP-hSOD1G85R overexpression induced SOD1 inclusions and reduced proteasome activity compared with the overexpression of GFP alone in NSC34 motor neuronal cells. In addition, we also observed that BV treatment restored proteasome activity and reduced the accumulation of ubiquitinated and misfolded SOD1 in GFP-hSOD1G85R-overexpressing NSC34 motor neuronal cells. However, BV treatment did not activate the autophagic pathway in these cells.
  
  
  CONCLUSIONS
  Our findings suggest that BV may rescue the impairment of the UPS in ALS models.
authors.forename: Seon Hwy
authors.initials: SH
authors.lastname: Kim
authors.forename: So Young
authors.initials: SY
authors.lastname: Jung
authors.forename: Kang-Woo
authors.initials: KW
authors.lastname: Lee
authors.forename: Sun Hwa
authors.initials: SH
authors.lastname: Lee
authors.forename: MuDan
authors.initials: M
authors.lastname: Cai
authors.forename: Sun-Mi
authors.initials: SM
authors.lastname: Choi
authors.forename: Eun Jin
authors.initials: EJ
authors.lastname: Yang
mesh_terms: D000690:Amyotrophic Lateral Sclerosis / Q000188:drug therapy / Q000378:metabolism*; D000818:Animals; D056705:Apitherapy; D001343:Autophagy; D001514:Bee Venoms / Q000494:pharmacology*; D001516:Bees; D000091342:Endoplasmic Reticulum Chaperone BiP; D006801:Humans; D051379:Mice; D009046:Motor Neurons / Q000187:drug effects* / Q000378:metabolism; D009154:Mutation; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism*; D013482:Superoxide Dismutase / Q000235:genetics / Q000378:metabolism*; D000072105:Superoxide Dismutase-1; D014162:Transfection; D025801:Ubiquitin / Q000378:metabolism*
chemical_list: D001514:Bee Venoms; D000091342:Endoplasmic Reticulum Chaperone BiP; C000717247:HSPA5 protein, human; C000717248:Hspa5 protein, mouse; C000606290:SOD1 protein, human; D025801:Ubiquitin; C000606291:Sod1 protein, mouse; D013482:Superoxide Dismutase; D000072105:Superoxide Dismutase-1; D046988:Proteasome Endopeptidase Complex