The Comparative Study Of BYHWD And XFZYD Combined With EPC To Repair Injured Vascular Endothelial

Objective:This study is based on traditional Chinese medicine theory of blood circulation and cell therapy new technical guidance.We researched Yiqihuoxue and xingqihuoxue promote endothelial progenitor cells to repair damaged vascular endothelium and its mechanism.BYHWD and XFZYD were selected to study both prescription joint EPC infusion repair the rat endothelial injury and Investigate the mechanism from homing and differentiation in the cellular and molecular level.Method:We investigate the BYHWD and XFZYD's recovery effect to injured Vascular endothelial in rat,after the two Chinese herbal compound united EPC infusion.Based on the clear synergies of the both united EPC,we study the mechanism of two Chinese herbal compound promote EPC repairing the damaged vascular endothelial from EPC differentiation and homing,and selected SDF-1/CXCR-4 axis,integrin molecule ?v?3,?v?5 and PI3K/Akt signaling pathway as starting point.Result:1.Get EPC with improved density gradient centrifugation,detect Cell surface markers CD34/CD133 on 7d?14d?21d.The results showed that the expression of anti-double were 16.9%,22.6%,19.71%.the EPC tags rate is up 95%using DAPI labeling.2.The SD rats show of Elevated serum lipids,vascular intimal thickening,and some vessels atherosclerosis-like changes,vascular endothelial cell senescence after continuous high-fat diet for three months,supplemented by intraperitoneal injection of vitamin D.Compared with group M,each experimental groups cellular senescence had improved,and group B,X and E improved especially after four weeks drug intervention�3.Compared with group M,group b?y?B?E could lower total cholesterol and triglycerides(P<0.05)and increase density lipoprotein(P<0.05);Compared with group E,group b?y?B?E could Lower total cholesterol and triglycerides(P<0.05);Compared with the group y,group B?E could Lower total cholesterol and triglycerides(P<0.05)and increase density lipoprotein(P<0.05);Compared with group M,each experimental groups Significantly lower serum calcium concentration(P<0.05);Compared with group E,group E decreased Calcium more significantly(P<0.05).Group b and group y compared with each other,group b is inferior to group y in the aspects of regulating triglycerides and High-density lipoprotein,but group B was superior to group X in the aspect of Regulation of blood lipids and calcium,especially lower total cholesterol,increase HDL and triglycerides,after two Chinese herbal compound united EPC(P<0.05).BYHWD has an additive effect in terms of total cholesterol regulation with EPC,Two Chinese herbal compound have a synergistic effect with transfusion alone EPC.4.Compared with group M,eNOS expression was significantly increased in each experimental groups,Wherein group B,X,E significantly enhanced(P<0.01).Compared with group E,eNOS expression was significantly increased in group B,X(P<0.05).Compared with group b and group y,group B and E could increase eNOS expression(P<0.05).BYHWD and XFZYD have an additive effect with EPC in terms of increasing eNOS expression.5.Compared with group M,group B SDF-1 protein expression increased significantly(p<0.01),group X SDF-1 protein expression increased(p<0.05);Compared with group E and group b?y,group B?E SDF-1 protein expression increased(p<0.05 or p<0.01);Compared with group C,group M and the rest of the experimental groups could decrease expression of CXCR4(p<0.05),compared with group M,there is no significant differences in the expression of CXCR4 in each experimental group.there is no significant difference between group b and group y,but group B could up-regulated SDF-1 protein significantly higher than group X after Combined with EPC.BYHWD has an synergistic effect with EPC in terms of up-regulating SDF-1 protein and XFZYD have an additive effect with EPC in this aspect.6.Compared with group M,the expression of ?v?3 increased in each group(p<0.05),and group B increased significantly(p<0.01),Compared with group E,the expression of ?v?3 decreased(p<0.05);Compared with group b,the expression of ?v?3 increased in group B(p<0.05).Compared with group M,the expression of ?v?5 increased in each experimental group,and there is no difference in each group(p>0.05).There is no difference between group b and group y,but group B was superior to group X in terms of up-regulating ?v?3 after Combined with EPC.BYHWD has an additive effect with EPC in terms of increasing ?v?3 expression.7.Compared with group M,the expression of PI3K enhanced in group B?X?b;compared with group b and group y,the expression of PI3K enhanced in group B?X(p<0.01 ? p<0.05);compared with group E,the expression of PI3K enhanced in group B?X(p<0.05).compared with group M,the expression of p-AKT enhanced in each experimental group(p<0.05orp<0.01);compared with group E and group b?y,the expression of p-AKT enhanced in group X(p<0.01).Group b was superior to group y in terms of up-regulating PI3K.BYHWD and XFZYD have an synergistic effect with EPC in terms of up-regulating PI3K and p-AKT.Conclusion:1.The improved density gradient centrifugation method could get EPC,and the best infusion time is primary culture of 14 day,DAPI labeled as long-term tracer EPC was feasibility.2.BYHWD and XFZYD could promote EPC significantly to repair the damaged vascular endothelial from Lipids,calcium detection,The repair of the vessel wall:fluorescence detection and the expression of eNOS.Comparison of the two parties BYHWD superior to XFZYD3.BYHWD and XFZYD could promote EPC homing to repair the damaged vascular endothelial through upregulation of SDF-1 and?v?3.BYHWD and XFZYD could promote EPC differentiation to repair the damaged vascular endothelial through upregulation PI3K/Akt signaling pathway.