title: Heparanase induced by advanced glycation end products (AGEs) promotes macrophage migration involving RAGE and PI3K/AKT pathway.
abstract: BACKGROUND
  Advanced glycation end products (AGEs), inflammatory-associated macrophage migration and accumulation are crucial for initiation and progression of diabetic vascular complication. Enzymatic activity of heparanase (HPA) is implicated strongly in dissemination of metastatic tumor cells and cells of the immune system. In addition, HPA enhances the phosphorylation of selected signaling molecules including AKT pathway independent of enzymatic activity. However, virtually nothing is presently known the role of HPA during macrophage migration exposed to AGEs involving signal pathway.
  
  
  METHODS
  These studies were carried out in Ana-1 macrophages. Macrophage viability was measured by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assays. HPA and AKT protein expression in macrophages are analysed by Western blotting and HPA mRNA expression by real time quantitative RT-PCR. Release of HPA was determined by ELISA. Macrophage migration was assessed by Transwell assays.
  
  
  RESULTS
  HPA protein and mRNA were found to be increased significantly in AGEs-treated macrophages. Pretreatment with anti-HPA antibody which recognizes the nonenzymatic terminal of HPA prevented AGEs-induced AKT phosphorylation and macrophage migration. LY294002 (PI3k/AKT inhibitor) inhibited AGEs-induced macrophage migration. Furthermore, pretreatment with anti-receptor for advanced glycation end products (RAGE) antibody attenuated AGEs-induced HPA expression, AKT phosphorylation and macrophage migration.
  
  
  CONCLUSIONS
  These data indicate that AGEs-induced macrophage migration is dependent on HPA involving RAGE-HPA-PI3K/AKT pathway. The nonenzymatic activity of HPA may play a key role in AGEs-induced macrophage migration associated with inflammation in diabetic vascular complication.
authors: Department of Nephrology, Shanghai Fifth People's Hospital, Fudan University, Shanghai 200240, China.
authors: Qiaojing
authors: Q
authors: Qin
authors: Jianying
authors: J
authors: Niu
authors: Zhaoxia
authors: Z
authors: Wang
authors: Wangjie
authors: W
authors: Xu
authors: Zhongdong
authors: Z
authors: Qiao
authors: Yong
authors: Y
authors: Gu
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D054442:Cell Migration Assays, Macrophage; D002465:Cell Movement / Q000276:immunology; D002470:Cell Survival; D003925:Diabetic Angiopathies / Q000276:immunology* / Q000378:metabolism; D005966:Glucuronidase / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D017127:Glycation End Products, Advanced / Q000276:immunology*; D007249:Inflammation; D008264:Macrophages / Q000276:immunology* / Q000378:metabolism; D051379:Mice; D019869:Phosphatidylinositol 3-Kinases / Q000276:immunology* / Q000378:metabolism; D010766:Phosphorylation; D051057:Proto-Oncogene Proteins c-akt / Q000276:immunology* / Q000378:metabolism; D000067759:Receptor for Advanced Glycation End Products; D011971:Receptors, Immunologic / Q000276:immunology* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000276:immunology; D000099175:Heparanase
chemical_list: D005966:Glucuronidase; D017127:Glycation End Products, Advanced; D019869:Phosphatidylinositol 3-Kinases; D051057:Proto-Oncogene Proteins c-akt; D000067759:Receptor for Advanced Glycation End Products; D011971:Receptors, Immunologic; D000099175:Heparanase