The Animal Experimental Study Of Traditional Chinese Medicine And Transplantation With Endothelial Progenitor Cells For Treating Arteriosclerosis Obliterans In Lower Extremity

Objective In order to support theoretical basis and experimental evidence for clinical application of traditional Chinese medicine(TCM) and stem cells transplantation,which treating arteriosclerosis obliterans in lower extremity.To observe the possibility and effects of angiogenesis by TCM and transplantation with endothelial progenitor cells(EPCs) from bone marrow treating the animal experimental models of arteriosclerosis obliterans in lower extremity.Methods The rats experimental models with both arteriosclerosis obliterans in lower extremity were successfully made by feeding high-fat diet, Clamping both femoral arteries,which branehes were ligated and excised in 36 male Wistar rats. On the 12 week after operation,test the total cholesterol(TC),triglyceride(TG),high-density lipoprotein(HDL) and low-density lipoprotein(LDL) of 6 rats,which were sacrificed and their femoral-oopliteal arteries were examined by histological examinations. Mesenchymal stem cells(MSCs) were separated from bone marrow of rats, with density gradient centrifugation, adherent culture and amplified in vitro.The EPCs were cultured in mediun supplemented with inducing factor. Culture cells were identified by flow cytometry(FCM) to analyze the expression of CD34,CD133 and FLK-1.The EPCs labled with BrdU were transplanted into right hindlimbs of 24 rats of experimental models by intramuscular injection,the same volume of normal saline were injected into the left hindlimbs.12 rats named Group B were continuously treated by TCM for 14 days,the others were named Group A.On the 28 day after treatment, test TC,TG,HDL and LDL.On the 14 day and 28 day after treatment, the rats were sacrificed respectively and their muscles of hindlimbs were extracted for immunohistochemieal examinations by Fâ…§and BrdU.Observe the neoformative blood capillary density of the two groups under microscope and count blood capillaries with positive staining of Fâ…§each visual field.The statistical data were treated by ANOVA to analysis changes.Results Treating by TCM and transplantation with EPCs for 28 days,the results of TC,TG,HDL and LDL in Group A and Group B were more obviously improved than that of those without treatment(P<0.05), the results of Group B were more significantly improved than that of Group A(P<0.05).After treating for 14 days and 28 days,the rats in the two groups were found angiogenesis in the muscles of hindlimbs.The average number of the blood capillaries with positive staining of Fâ…§each visual field in Group B was more obviously than that in Group A(P<0.05).In the Group A and Group B,some positive stained EPCs by BrdU were found in the muscles of right hindlimbs not in the left hindlimbs.Conclusion EPCs can be isolated from MSCs and ampflified in vitro.Transplantation of EPCs and treatment of TCM can significantly increase blood capillary density and participate angiogenesis to improve blood supply and regulate serum lipid levels of rats with ischemic hindlimbs.