The Role And Mechanism Of Basic Fibroblast Growth Factor In Hypoxia Induced Rat Pulmonary Artery Smooth Muscle Cell Proliferation

Objective:To explore whether basic fibroblast growth factor(bFGF)participate in hypoxia induced rat pulmonary artery smooth muscle cells(PASMCs)proliferation and the relationship with cyclinD1.Methods:The healthy male SPF grade rats,rat PASMCs were isolated and cultured,and spread to three generations,PASMCs respective in hypoxia and normoxia,Hypoxia group is divided into 10%,5%,2%,1% oxygen concentration,respectively,to cultivate PASMCs 24 h,48 h,72 h,observe the cell growth situation,determined by MTT method to detect different oxygen concentration PASMCs proliferation,the most active oxygen concentration selection breeding as a follow-up experiments in the oxygen concentration of hypoxia groups.Cells can be divided into four groups by random number method: 1?normoxia group,2 ?hypoxia group,3?hypoxia+ NC siRNA group,4?hypoxia+ bFGF siRNA group,5,hypoxia + cyclinD1 siRNA group,respectively,to cultivate the rat PASMCs 24 h,48 h,72 h,the expression of bFGF,cyclinD1 mRNA and protein in different culture conditions,were detection by RT-PCR,Western blot.Results:1?In vitro cultured rat PASMCs is confirmed by morphology and ?-actin immune cell chemical dyeing.2?Hypoxia induced rat PASMCs proliferation,through depicting the growth curve and determined by MTT method found that in 1% oxygen concentration PASMCs proliferation are the most active.3?Compared with normoxia,indicated that hypoxia enhanced the expression of bFGF and cyclinD1 mRNA andproteins.4? The levels of bFGF and cyclinD1 mRNA and proteins was inhibited,when rat PASMCs exposed to hypoxia transfected with bFGF siRNA.when rat PASMCs exposed to hypoxia transfected with cyclinD1 siRNA,the levels of cyclinD1 mRNA and proteins was inhibited,while bFGF mRNA and protrin had no effect.Conclusion: 1?Hypoxia induced rat PASMCs poliferation,and hypoxia upregulated bFGF levels in PASMCs.2?Upregulation of bFGF is associated with hypoxia induced PASMCs poliferation via upregulation the expression of cyclinD1.