title: [Influence of high glucose and mannose binding lectin complement pathway activation to IL-6 and TNF-alpha's expression by human renal glomerular endothelial cells].
abstract: OBJECTIVE
  To investigate the effect of high glucose and mannose binding lectin (MBL) complement pathway activation's effect on expression of Interleukin-6 (IL-6) and Tumor necrosis factor-alpha (TNF-alpha) from human renal glomerular endothelial cells (HRGEC), to explore unknown pathogenesy of diabetic nephropathy.
  
  
  METHODS
  Normal HRGEC was divided randomly into normal glucose group(5 mmol/L D-glucose), manicol group (5 mmol/L D-glucose+25 mmol/L manicol) and high glucose group (30 mmol/L D-glucose). Real-time PCR was used to detect IL-6 and TNF-alpha mRNA expression in each group, Euzymelinked Immunosorbent Assay (ELISA) was performed to examine the protein expression of IL-6 and TNF-alpha in supernatant after 24 hours' culture. HRGEC was then randomly divided into two groups: single high glucose group and high glucose + MBL group. After 24 hours' culture with 30 mmol/L D-glucose, 30% MBL deficiency human serum was added into two groups, 1 microg/mL MBL was only added into high glucose + MBL group, continued the culturation for another 4 hours. Flow cytometry and immunofluorescence technique were applied to evaluate MBL, C3 and membrane attacks complex (MAC) deposition on cell surface respectively. Real-time PCR and ELISA were performed to examine mRNA and protein expression of both IL-6 and TNF-alpha in each group.
  
  
  RESULTS
  Compared with normal glucose group and manicol group, the mRNA and protein expression of IL-6 and TNF-alpha in high glucose group were increased (P < 0.05). Flow cytometry confirmed obvious MBL and C3 co-deposition and Immunofluorescence confirmed obvious MAC deposition on cell surface in high glucose+ MBL group. Compared with single high glucose group, the mRNA and protein expression of IL-6 and TNF-alpha in high glucose+ MBL group were significantly higher (P < 0.05).
  
  
  CONCLUSIONS
  High glucose can bring inflammatory factors' overexpression from cultured HRGEC; high glucose together with MBL can bring MBL complement pathway activation and inflammatory factors' overexpression, this indicates that the activation of MBL complement pathway may be a potential unknown pathogenesy of diabetic nephropathy and its proinflammatory status.
authors.affiliation: Department of Nephrology, West China Hospital, Sichuan University, Chengdu 610041, China.
authors.forename: Xiao-hui
authors.initials: XH
authors.lastname: Wu
authors.forename: Song-min
authors.initials: SM
authors.lastname: Huang
authors.forename: Wen-xing
authors.initials: WX
authors.lastname: Fan
authors.forename: Wan-xin
authors.initials: WX
authors.lastname: Tang
authors.forename: Hong-yu
authors.initials: HY
authors.lastname: Qiu
mesh_terms: D002478:Cells, Cultured; D037582:Complement Pathway, Mannose-Binding Lectin / Q000502:physiology*; D003928:Diabetic Nephropathies / Q000209:etiology; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism; D005947:Glucose / Q000494:pharmacology*; D006801:Humans; D015850:Interleukin-6 / Q000235:genetics / Q000378:metabolism*; D007678:Kidney Glomerulus / Q000166:cytology* / Q000378:metabolism; D037601:Mannose-Binding Lectin / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000378:metabolism*
chemical_list: D015850:Interleukin-6; D037601:Mannose-Binding Lectin; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; D005947:Glucose