PMID- 16620701 OWN - NLM STAT- MEDLINE DCOM- 20060525 LR - 20081121 IS - 0366-6999 (Print) IS - 0366-6999 (Linking) VI - 119 IP - 7 DP - 2006 Apr 5 TI - Brain-derived neurotrophic factor inducing angiogenesis through modulation of matrix-degrading proteases. PG - 589-95 AB - BACKGROUND: Recent studies have proved that brain-derived neurotrophic factor (BDNF) possesses angiogenic activity in vitro and in vivo. However, the proangiogenic mechanism of BDNF has not yet been provided with enough information. To explore the proangiogenic mechanism of BDNF, we investigated the effects of BDNF on extracellular proteolytic enzymes, including matrix metalloproteinases (MMPs) and serine proteases, particularly the urokinase-type plasminogen activator (uPA)-plasmin system in human umbilical vein endothelial cells (HUVECs) model. METHODS: Tube formation assay was performed in vitro to evaluate the effects of BDNF on angiogenesis. The HUVECs were treated with various concentrations of BDNF (25 - 400 ng/ml) for different (6 - 48 hours), reverse transcriptase-polymerase chain reaction (RT-PCR) was used to assay MMP-2, MMP-9, TIMP-1, and TIMP-2 mRNA in HUVECs, and the conditioned medium was analyzed for MMP and uPA activity by gelatin zymography and fibrin zymography, respectively. uPA, plasminogen activator inhibitor (PAI)-1, tissue inhibitors of metalloproteinase (TIMP)-1, and TIMP-2 were quantified by western blotting analysis. RESULTS: BDNF elicited robust and elongated angiogeneis in two-dimensional cultures of HUVECs in comparison with control. The stimulation of serum-starved HUVECs with BDNF caused obvious increase in MMP-2 and MMP-9 mRNA expression and induced the pro-MMP-2 and pro-MMP-9 activation without significant differences in proliferation. However, BDNF had no effect on TIMP-1 and TIMP-2 production. BDNF increased uPA and PAI-1 production in a dose-dependent manner. Maximal activation of uPA and PAI-1 expression in HUVECs was induced by 100 ng/ml BDNF, while effects of 200 ng/ml and 400 ng/ml BDNF were slightly reduced in comparison with with those of 100 ng/ml. Protease activity for uPA was also increased by BDNF in a dose-dependent manner. BDNF also stimulated uPA and PAI-1 production beyond that in control cultures in a time-dependent manner from 12 hours to 48 hours after BDNF treatment. CONCLUSIONS: BDNF stimulates MMP and uPA/PAI-1 proteolytic network in HUVECs, which may be important to the acquisition of proangiogenic potential. FAU - Sun, Chun-yan AU - Sun CY AD - Institute of Hematology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China. FAU - Hu, Yu AU - Hu Y FAU - Wang, Hua-fang AU - Wang HF FAU - He, Wen-juan AU - He WJ FAU - Wang, Ya-dan AU - Wang YD FAU - Wu, Tao AU - Wu T LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't PL - China TA - Chin Med J (Engl) JT - Chinese medical journal JID - 7513795 RN - 0 (Brain-Derived Neurotrophic Factor) RN - 0 (Plasminogen Activator Inhibitor 1) RN - 0 (RNA, Messenger) RN - EC 3.4.21.73 (Urokinase-Type Plasminogen Activator) RN - EC 3.4.24.24 (Matrix Metalloproteinase 2) RN - EC 3.4.24.35 (Matrix Metalloproteinase 9) SB - IM MH - Brain-Derived Neurotrophic Factor/*pharmacology MH - Cells, Cultured MH - Gene Expression Regulation/*drug effects MH - Humans MH - Matrix Metalloproteinase 2/*genetics MH - Matrix Metalloproteinase 9/*genetics MH - Neovascularization, Physiologic/*drug effects MH - Plasminogen Activator Inhibitor 1/genetics MH - RNA, Messenger/analysis MH - Urokinase-Type Plasminogen Activator/genetics EDAT- 2006/04/20 09:00 MHDA- 2006/05/26 09:00 CRDT- 2006/04/20 09:00 PHST- 2006/04/20 09:00 [pubmed] PHST- 2006/05/26 09:00 [medline] PHST- 2006/04/20 09:00 [entrez] PST - ppublish SO - Chin Med J (Engl). 2006 Apr 5;119(7):589-95.