| Objective:To investigate the effect of Danshen Tongluo Jiedu decoction(DSTLJD)combined with endothelial progenitor cells(EPCs)transp-lantation on angiogenesis in rats with myocardial ischemia reper-fusion injury(IRI)and its mechanism.Methods:Using density gradient centrifugation and differential adherence method to isolate and culture EPCs,identification of EPCs by immunofluorescence.The IRI model was established by ligating the left anterior descending coronary artery,Grouping by means of random digital tables:sham operation group,IRI group,EPCs transplantation gloup,DSTLJD gloup,DSTLJD combined with EPCs transplantation gloup,PI3K inhibitor group,10 rats in each group.EPCs transplantation group,DSTLJD combined with EPCs transplantation group,PI3K inhibitor group were injected with EPCs suspension into the tail vein of rats,and DSTLJD gloup,DSTLJD combined with EPCs transplantation gloup,PI3K inhibitor group treated with DSTLJD on the 1 day after operation,twice a day for gastric lavage,the remaining group were given normal saline to fill the stomach.PI3K inhibitor group were injected with LY294002 into the tail vein of rats.After 4 weeks,the pathological morphology of myocardium was observed by hematoxylin eosin staining,to detect the expression of CD34 in myocardial tissue of rats by immunohistochemical method,the myocardial microvessel count(MVC)and myocardial microvascular density(MVD)was calculated and compared,the expression of vascular endothelial growth factor(VEGF)and basic fibroblast growth factor(bFGF)in each group were detected.Detection of the protein expression of PI3K,p-AKT and AKT in rats of each group by western blot.Results:Compared with IRI group,the degree of myocardial injury was improved in EPCs transplantation group,DSTLJD group,DSTLJD combined with EPCs transplantation gloup,these three groups with MVC,MVD,VEGF and bFGF protein expression,PI3K and p-AKT were significantly increased,the difference was statistically significant(P<0.01),the DSTLJD group was superior to the EPCs transplantation group,and the difference was statistically significant(P<0.05).Compared with EPCs transplantation group and DSTLJD group,the degree of myocardial injury was obviously improved in DSTLJD combined with EPCs transplantation gloup,the gloup with MVC,MVD,VEGF and bFGF protein expression,PI3K and p-AKT were significantly increased,the difference was statistically significant(P<0.01).Compared with DSTLJD combined with EPCs transplantation gloup,on the basis of Danshen Tongluo Jiedu decoction combined with endothelial progenitor cell transplantation,after application of PI3K inhibitor,the degree of myocardial injury in the PI3K inhibitor group was more severe,the gloup with MVC,MVD,VEGF and bFGF protein expression,there was no significant difference in the expression of PI3K(P>0.05),p-AKT were significant reduction,the difference was statistically significant(P<0.01).Conclusion:1.Danshen Tongluo Jiedu decoction can obviously reduce the infiltration of inflammatory cells and improve the pathological morphology of myocar-dium.2.Danshen Tongluo Jiedu decoction could significantly increase MVC and MVD,it can upregulate the expression of VEGF and bFGF,it increase the expression of PI3K and p-AKT,it promoting angiogenesis in IRI rats,the mechanism may be related to the regulation of PI3K/AKT signaling pathway.3.Danshen Tongluo Jiedu decoction combined with endothelial progenitor cell transplantation can obviously improve the pathological morphology of myocardial tissue,it could significantly increase MVC and MVD,it can upregulate the expression of VEGF and bFGF,it increase the expression of PI3K and p-AKT,it promoting angiogenesis in IRI rats,so as to protect the myocardium of IRI rats,the mechanism may be related to the regulation of PI3K/AKT signaling pathway. |
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