Study On Human Endothelial Cell αv Integrin And MMP-1 Stimulated By Basic Fibroblast Growth Factor

ObjectiveTrauma is one of the dangerous diseases which occur widely in advanced society. So the problem of the wound healing is still the emphasis in medical research. The wound healing is a kind of complex course involving many factors and cells. Among them the effects of growth factors have become the focus of the research . Fibroblast growth factors ( FGF) especially the basic fibroblast growth factors (bFGF) have extensively biological activity , which could influence every aspects in repair. Thus it is important to study its mechanism deeply for the applying of FGF in clinics. While so for , the report about this is quite a few. In the current study to explore the possibe mechanism of endothelial cell (EC) migration and angiogenesis by bFGF, the 'expression of av integrin and secretion of MMP - 1 were detected on human umbilical vein endothelial cell ( HUVEC) which were stimulated by bFGF,as well as the effect of bFGF on TNP -470.MethodsAt first the HUVECs were cultured. The serum - free media was used as the cells were sub - confluent. To detect av integrin ; the HUVEC were divided into 4 groups : control (serum -free) ; bFGF(500ng/ml) ; TNP-470(10-7g/ ml) ; bFGF + TNP -470.72hours later the expression was detected by immuno-cytochemistry and flow cytometry. To detect MMP - 1: control( serum - free) ; bFGF(300ng/ml,500ng/ml,700ng/ml) ,24hours later the secretion of MMP -1 was detected by western blotting.ResultsAccording to the analysis of immunocytochemistry and flow cytometry bFCF increase markedly the expression of HUVEC av integrin, TNP - 470 decreases the expression of av integrin. bFGF can inverse the inhibition of TNP - 470 ( p <0.05). the results of western blotting ;the secretion of MMP -1 of EC was increased by bFGF (p <0.05). the data was analyzed by SPSS.DiscussionIt' s proved by many experimens that bFGF could promot migration of EC and angiogenesis during wound healing. However the associated mechanisms were unclear. To explore the possible mechanism of bFGF, We tested the expression of av integrin and secretion of MMP - 1 on EC which was stimulated by bFGF.av integrin could interact with extracellular matrix as a transmembrane gly-coprotein on EC. Tlius promoting EC migrating in ECM. According to our experimental results we find that the bFGF could increase the expression ofav integrin on EC. So we guess that bFGF maybe promote EC migrating and angiogenesis due to upregulating the expression of av integrin.TNP-470 is a specific antiangiogenic agent, which could inhibit the migration of EC. From our experiments we know that TNP -470 decrease the expression of av integrin on EC, while bFGF could reverse this inhibition.Matrix metalloproteinase - 1 ( MMP - 1) is interstitial collagenase which is correlative with angiogenesis during wound healing. MMP - 1 could combine with I collagen and degrade it. in addition, MMP -1 can degrade collagen II III VIII X ,then damage the barriers of migration for EC. From our experimental results we find that bFGF promots EC secretin MMP -1 so we infer that bFGF enhance migration of EC which is highly correlative with the augment of MMP -1 secretion by bFGF.Conclusion1. bFGF promotes the expression of HUVEC αv integrin.2. bFGF enhances the secretion of HUVEC MMP -1.3. TNP-470 decreases the expression of HUVEC αv integrin,bFGF can inverse the inhibition of TNP -470.