The Role Of Soluble Vascular Endothelial Growth Factor Receptor Flt-1(sFlt-1) In Atherosclerotic Plaque

Objective: To construct the soluble vascular endothelial growth factor receptor Flt-1 (sFlt-1) eukaryotic expression vector pEGFP-N1-sFlt-1 and transfect it into iliac artery of rabbit. Detecting the exppression of sFlt-1 in HUVEC and observing the effects of high-expressed sFlt-1 on atherosclerotic plaque.Method:1.Construction of the sflt-1 cDNA reconbination gene eukaryotic expression vector:Obtained the target gene with the method of enzyme digestion from clone vector. Enzyme digested target vector and purificated target production, linked each other and transformed it into competence cells. Identified positive clones by PCR.The identified positive clones were proved that it had been linked with the target vector . sequenced and contrasted the positive clones.The correct sequence was the fusion protein expression plasmid vector. 2. Cultivation of HUVEC: The pEGFP-N1-sFlt-1 recombination plasmid was transfected into cultured HUVEC in vitro. Detected the expression of sFlt-1 gene with fluorescence and western blotting. 3.The effects of sflt-1 gene on atherosclerotic plaque: 48 male New Zealand rabbits were divided into 4 groups randomly.â‘ Normal control group(n=8).â‘¡Sham operation group(n=8).â‘¢Balloon injury+high lipid diet+ pEGFP-N1 group(n=16).â‘£Balloon injury+high lipid diet+ pEGFP-N1-sFlt-1 group(n=16).Groupâ‘ and groupâ‘¡were supplied with normal diet. Groupâ‘¡was suffered with sham operation.Groupâ‘¢and groupâ‘£were supplied with high lipid diet and injuried by balloon. Groupâ‘¢was transfected with pEGFP-N1 and groupâ‘£transfected with pEGFP-N1-sFlt-1.Detected the local expression of sFlt-1 by western blotting after three days. Detected blood lipid at the beginning and the end of the experiment.Measured the morphology of atherosclerotic plaque by HE stain and the formation of newly formed vessels by CD34 positive cells of immunohistochemistry.Results:1. The sFlt-1 recombination plasmid was successfully constructed. 2. sFlt-1 expressed effectively after transfected into HUVEC. 3. The model of rabbits iliac artery atherosclerotic plaque was successfully established.4.sFlt-1 inhibited the formation and development of atherosclerotic plaque.Conclusion:The local high-expressed sFlt-1 can inhibit the formation and development of atherosclerotic plaque.