BMSCs transplantation of Danshen injection to treatment of limb ischemia to promote angiogenesis

Objective: To investigate the effect of autologus bone marrow mesenchymal stem cell(BMSCs) transplantation and its combination with Danshen injection for the New Zealand rabbit limb ischemia model.Methods: 18 New Zealand rabbits,through the thread ligation of vessels resulting in limb ischemia models,were divided into 3 groups: the control group,namely the physiological saline group(NS);the BMSCs transplantation group,the autologous bone marrow mesenchymal stem cell transplantation group(BMSC);the combined treatment group,the autologous bone marrow mesenchymal stem cell transplantation combined with Salvia miliorrhiza injection group(BMSCs+D).A week after the model was established,in the BMSCs transplantation group and the combination group,autologous BMSCs cultured were injected into the gastrocnemius muscle subcutaneously at multipoints,the control group was treated with saline and the combination group was treated with auricular intravenous drip of Danshen on the basis of BMSCs transplantation.After two weeks of conventional breeding,(1) ultrasonic tests were conducted of angiogenesis indicators,superficial femoral artery diameters,blood flow signals,peak velocity and acceleration time,(2) by immunohistochemical staining,Brdu staining positive cells were observed and compared in all the groups and(3) through HE staining angiogenesis was observed and compared.Results:(1) Ultrasound findings: ①2D ultrasound showed,in the combination group and the BMSCs transplantation group,femoral artery and proximal superficial femoral artery diameters widened significantly compared with the control group(P0.05),and in the combination therapy group,the diameters widened more significantly than in the BMSCs transplantation group(P0.05);② CDFI ultrasound showed,in the combination group and the BMSCs transplantation group,the femoral artery blood flow signals were enhanced more obviously than in the control group and the signals were enhanced more obviously in the combination group than in the BMSCs transplantation group;③ the colour doppler images revelaed that in the BMSCs transplantation group and the combination group,the femoral artery blood flow peak velocity increased compared with the control group,and the blood flow acceleration time was significantly less than that in the control group(P0.05)and the acceleration time was slightly greater in the combination group than in the BMSCs transplantation group(P0.05).(2) Immunohistochemical staining: anti Brdu staining positive cells were detected in the BMSCs transplantation group and the combination group and more cells were found in the combination group than in the BMSCs transplantation group.(3) HE staining: the vascular density in the BMSCs transplantation group and the combination group was significantly higher than in the control group and it was higher in the combination group than in the BMSCs transplantation group.Conclusion: the indicators for angiogenesis in the BMSCs transplantation group and the combination group is obviously superior to those in the control group,and they are better in the combination group than in the BMSCs transplantation group.