title: [Protective effect of adipocytes fatty acid binding protein inhibitor on pancreas islet cell against macrophage-mediated cytotoxicity of mice in vitro].
abstract: OBJECTIVE
  To investigate the potential role of adipocytes fatty acid binding protein (A-FABP)inhibitor to prevent pancreatic islet cells from cytotoxic injury by inflammatory cytokines released from macrophage.
  
  
  METHODS
  Co-culture system for RAW264.7 macrophage and MIN6 insulinoma cells was established through transwell combined with A-FABP inhibitor BMS309403 treatment for 48 h. Meanwhile, cultured RAW264.7 and MIN6 respectively were set up as controls. In the inhibitor group, BMS309403 preprocessing (5 µmol/L) was performed 2 h before co-culture. The expression of toll-like receptors(TLR)4 and A-FABP in RAW264.7 macrophages was detected by RT-PCR and Western blotting, interleukin (IL)-1β and tumor necrosis factor(TNF)-α levels in the supernatant were detected by ELISA, Glucose-stimulated insulin level was detected by insulin radioimmuno-assay kits for the function of islets.
  
  
  RESULTS
  (1) The mRNA and protein levels of both TLR4 and A-FABP in RAW264.7 macrophages as well as the concentrations of IL-1β and TNF-α in the supernatant were significantly higher in co-culture group than in macrophages control group (P < 0.05). (2) Insulin secretion stimulated by high glucose was obviously decreased in co-culture group when compared with insulinoma cells control group [(16.0 ± 2.2) vs (41.1 ± 6.6) ng/ml, P < 0.05]. After the treatment with A-FABP inhibitor, the mRNA and protein levels of both TLR4 and A-FABP as well as the concentrations of IL-1β and TNF-α in the supernatant were significantly lower than in co-culture control (P < 0.05). However, insulin secretion stimulated by high glucose was significantly enhanced when compared with insulinoma cells control group [(31.4 ± 3.3) vs (16.0 ± 2.2) ng/ml, P < 0.05].
  
  
  CONCLUSIONS
  This study demonstrated that co-culture of macrophage and islet cells can activate inflammation pathway, stimulate inflammatory cytokine release and decrease insulin secretion from islet cells. A-FABP inhibitor can protect islet cells from macrophage-mediated cytotoxicity and preserve its insulin secretory function.
authors: Institute of Metabolism and Endocrinology, the Second Xiangya Hospital Diabetes Center, Central South University, Key Laboratory of Diabetes Immunology, Ministry of Education, Changsha 410011, China.
authors: Shi-ping
authors: SP
authors: Liu
authors: Li-ning
authors: LN
authors: He
authors: Yang
authors: Y
authors: Xiao
authors: Xin-ying
authors: XY
authors: Li
authors: Zhi-guang
authors: ZG
authors: Zhou
mesh_terms: D000818:Animals; D002460:Cell Line; D018920:Coculture Techniques; D050556:Fatty Acid-Binding Proteins / Q000037:antagonists & inhibitors*; D007249:Inflammation; D007328:Insulin / Q000378:metabolism; D000078790:Insulin Secretion; D053583:Interleukin-1beta / Q000032:analysis; D007515:Islets of Langerhans / Q000187:drug effects* / Q000378:metabolism; D008264:Macrophages / Q000378:metabolism*; D051379:Mice; D051197:Toll-Like Receptor 4 / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000032:analysis
chemical_list: D050556:Fatty Acid-Binding Proteins; D007328:Insulin; D053583:Interleukin-1beta; C493487:Tlr4 protein, mouse; D051197:Toll-Like Receptor 4; D014409:Tumor Necrosis Factor-alpha