The Expriment Research On Effect Of Pigment Epithelium-derived Factor On Proliferation And Apoptosis Of Vascular Smooth Muscle Cells After Hypoxia-reoxygenation

BackgroundOur country has more than four million the myocardial infarction (MI) pa-tients, And every year has at least five hundred thousand at the rate of growth,This disease is a serious threat to people’s life and health, At present, the surgicaltherapy of traditional to coronary artery stenosis or myocardial cell diffuse largenecrotic has been formed a large scar of poor patients is not effect, In recentyears, the stem cell transplantation for the treatment of the MI brings new hopefor these patients. In many of the stem cells, bone marrow stromal stem cells(MSCs) have are biological characteristics which are easy to isolate, culture,amplify, the low immunogenicity and are easy chemotactic to the damaged partand so on, become the ideal transplantation cells, Which is the international re- search hot spot. More and more studies show that MSCs can promote the recov-ery of MI mainly through their paracrine effect. in part infarction, MSCs cansecrete plenty of active factors that may play a variety of biological action.PEDF is an important cell factor that MSCs secrete, which belong to serpinprotease inhibitors superfamily member without protease inhibitors of function.PEDF widely exists in the human organization, With neurotrophic and neuro-protective, antitumorigenic, antiangiogenic, antioxidation, anti-inflammatoryand antithrombotic and other characteristics. Among them, antiangiogenic func-tion caused widely attention, But the mechanism of antiangiogenic is not en-tirely clear. Previous studies have focused on biological actions of angiogenesisfactor that MSCs secrete to MI pathological process and mechanism research,But antiangiogenic factor research is scare, This research through Hy-poxia-reoxygenation (H/R) model simulation ischemia-reperfusion injury, Pre-liminary detect the effect of recombinant human epithelium-derived factor(PEDF) on vascular smooth muscle cells (VSMCs) proliferation and apoptosisand its mechanism after hypoxia／reoxygen (H/R), For MSCs to treat MI bringsnew targets.MethodsVSMCs were primarily cultured by tissue-sticking method and were ran-domly divided into three groups, Respectively, Control group, H/R model groupand model+PEDF group. According to the concentration of PEDF, themodel+PEDF group was sub-divided into50ng/ml,100ng/ml,200ng/ml and400ng/ml groups. VSMCs proliferation was determined by MTT and cell apop-tosis was detected by Hoechst/PI staining and flow cytometry. The expressionsof Bcl-2and Bax protein were tested by Western blot． Results1．MTT showed that H/R increased VSMCs proliferation and PEDF inhib-ited the pro1iferation of VSMCs. The inhibitive effect of200ng/ml PEDF wasthe most significant (P <0.01).2．After VSMCs were treated with200ng/ml PEDF+H/R, Hoechst/PIstaining and flow cytometry demonstrated that the apoptosis rate of the cellssignificantly increased(P <0.01vs control group and H/R).3．Western blot demonstrated that VSMCs were treated with200ng/mlPEDF+H/R could down-regulated Bcl-2expression and up-regulated Bax pro-tein expression (P <0.01vs control group and H/R).Conclusion1．H/R induces VSMCs abnormal proliferation and PEDF inhibits VSMCsproliferation, The inhibitive effect of200ng/ml PEDF was the most significant.But PEDF can not inhibit VSMCs proliferation without H/R, which make PEDFcomprehensive used in clinic.2．PEDF induces VSMCs apoptosis after H/R possibly through up-regulatedBax protein expression and down-regulated Bcl-2expression. It will be furthercomfirmed whether there are other pathways.3．VSMCs were primarily cultured by tissue-sticking method, which arehigh purity, grow flourish, operate simply, high success rate.