title: [Effect of metabolic drugs on the secretion of acylation stimulating protein in 3T3-L1 adipocytes].
abstract: OBJECTIVE
  To develop a 3T3-L1 pre-adipocyte model for evaluating the secretion of acylation stimulating protein (ASP) and an ELISA assay for measuring ASP production and investigate the effects and related potential mechanisms of metabolic drugs on the secretion of ASP,  and on the complement C3,  triglyceride (TG) mass,  nonesterified fatty acids (NEFA) release and fatty acid (FA) uptake into adipocytes.
  
  
  METHODS
  After differentiated,  3T3-L1 pre-adipocytes were treated with chylomicrons,  metformin,  rosiglitazone,  rimonabant for 48 h. ASP and C3 were measured using a sandwich ELISA. NEFA levels were measured using enzymatic colorimetric kits. FA uptake was measured in a bottom-reading fluorescent microplate reader. TG mass and protein levels were determined using enzymatic colorimetric assay and the Bradford assay,  respectively.
  
  
  RESULTS
  Chylomicrons increased ASP production (up to 411%±133%,  P<0.05). Rosiglitazone and rimonabant decreased ASP production (-53% to -85%,  P<0.05),  associated with a decrease in the precursor protein C3 (-37% to -65%,  P<0.01). By contrast,  metformin also decreased ASP (-54% to  -100%,  P<0.05),  but with no change in precursor protein C3. In addition,  metformin decreased TG mass (maximum -60%,  P<0.05) and real-time FA uptake (maximum -75%,  P<0.05).
  
  
  CONCLUSIONS
  3T3-L1 pre-adipocyte model is successfully developed,  which can be used for evaluating the effects of metabolic drugs on the secretion of ASP.  ELISA assay for measuring ASP production is also developed.
authors.affiliation: Department of Cardiology, Xinjiang Medical University, Urumqi, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Gao
authors.forename: Katherine
authors.initials: K
authors.lastname: Cianflone
authors.forename: Danny
authors.initials: D
authors.lastname: Gauvreau
authors.forename: Wei
authors.initials: W
authors.lastname: Cui
authors.forename: Yi-tong
authors.initials: YT
authors.lastname: Ma
mesh_terms: D041721:3T3-L1 Cells; D017667:Adipocytes / Q000187:drug effects* / Q000378:metabolism*; D000818:Animals; D002914:Chylomicrons / Q000378:metabolism / Q000494:pharmacology; D003176:Complement C3 / Q000378:metabolism; D005230:Fatty Acids, Nonesterified / Q000378:metabolism; D007004:Hypoglycemic Agents / Q000494:pharmacology; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism*; D051379:Mice; D010880:Piperidines / Q000494:pharmacology; D011720:Pyrazoles / Q000494:pharmacology; D000077285:Rimonabant; D014280:Triglycerides / Q000378:metabolism
chemical_list: C545610:C3 protein, mouse; D002914:Chylomicrons; D003176:Complement C3; D005230:Fatty Acids, Nonesterified; D007004:Hypoglycemic Agents; D036341:Intercellular Signaling Peptides and Proteins; D010880:Piperidines; D011720:Pyrazoles; D014280:Triglycerides; D000077285:Rimonabant