The Effect Of BFGF On Inducing Mouse Cardiac Microvascular Endothelial Cell To Form New Blood Vessels

Therapeutic angiogenesis makes use of the administration of angiogenic growth factor protein or gene to promote the development of endogenous collateral vessels in ischemic heart disease. The goal of therapeutic angiogenesis is to promote the development of supplemental blood conduits that will act as endogenous bypass vessels. Among the growth factors that play a role in blood vessel growth and development, VEGF and bFGF have been the most extensively studied. Thus the experiment cultured cardiac micro vascular endothelial cell (CMEC) in vitro, and set up a stable angiogenesis culture system by Matrigel in order to discuss the influence of bFGF on inducing CMEC to form new blood vessels and the role of Fit - 1.MethodsFirstly, CMEC was cultured by modified Nishida method and identified. Divided into five groups;normal control ( serum - free DMEM culture liquid) and bFGF stimulating groups in varying concentrations (5,10,20,40ng/ml) , acting 24h. CMEC was induced to develop tube structure by using Matrigel so that to detect the influence of bFGF on neo'vascularization. RT - PCR assay the expression of Fit - 1mRNA. Finally, analysis experiment results.ResultsThe experiment results had shown that, CMEC can be induced to develop tube structure by Matrigel in vitro. After treatment with bFGF in varying coneen-trations (5,10,20,40ng/ml) ,the number of tubes higher than control group, and in a dose - dependent manner (in 0 - 20 ng/ml). Furthermore, after treatment with bFGF in varying concentrations (5 ,10,20,40ng/ml)for 24h, the expression of Fit — 1 mRNA higher than control group, it parallel to the development of tube structure by Matrigel.DiscussionTherapeutic angiogenesis, in the form of growth factor protein administration or gene therapy has emerged as a new method of treatment for patients with severe, inoperable ischemic heart disease. New vessel formation occurs through the processes of angiogenesis, vasculogenesis, and arteriogenesis, under the control of growth factors such as those that belong to the VEGF, bFGF.bFGF as a member of the FGF family, is released by most cell types. It induces endothelial cell proliferation, migration, and formation of new vessels. The research results also show that CMEC can be induced to develop tube structure by Matrigel.Additional, bFGF also improve the expression of VEGF and its receptors. VEGF family currently comprises six members ?. VEGF - A ^ VEGF - B ^VEGF -C NVEGF - D ,VEGF - E and placenta growth factor ( PIGF). VEGF and its high - affinity binding receptors, the tyrosine kinase Flk - 1 and Fit - 1, are thought to be important for the development of vasculature( endothelial cell proliferation , migration, basement membrane degradation and formation of new vessels). There are lots of reports about the influence of bFGF on VEGFR -2 and few reports about the influence of bFGF on VEGFR - 1. The Fit - 1 signaling pathway may be important in controlling blood vessel growth and development. The research results also show that bFGF can inhance the expression of Fit — lmRNA. It is a cue that bFGF influence neovascularization through VEGFR.Conclusion1. cardiac microvascular endothelial cell can be induced to develop tubestructure by Matrigel in vitro.2. bFGF can promote cardiac microvascular endothelial cell to form new blood vessels and in certain rage showing a dose - dependent manner3. bFGF can enhance the expression of Fit - 1 and in certain rage showing a dose - dependent manner , this dose - dependent manner similar to that of new blood vessels' quantity.