Endothelial Progenitor Cells Induced By Neurotrophins-3 Improved Treatments On Chronic Lower Limb Ischemia In Rats

Objective: Our research focuses on identifying the role of neurotrophins-3(NT-3)on proliferation,differentiation and function of endothelial progenitor cells derived from bone marrow(BM-EPCs),we also check the effect of NT-3 treated endothelial progenitor cells(EPCs)on lower limb ischemia in rats.Methods: First,we isolated early BM-EPCs from bone marrow of SD-rats through gradient centrifugation,using lymphocyte separation medium and obtained late BM-EPCs by collecting non-adherent early EPCs after 72 hours.In addition,we also isolated neurons from brain of neonatal SD-rats,using enzyme and mechanical methods.Second,we did BrdU incorporation,immunochemistry and other assays to check whether NT-3 conditional medium could accelerate proliferative rates,improve differentiate capacity and promote the ability to secrete vascular endothelial growth factor 165(VEGF165)of EPCs,and found out the best concentrations of NT-3 is 200ng/ml.Finally,we verified the role of NT-3 treated EPCs on treating lower limb ischemia in rats.Results:1.We isolated pure BM-EPCs and neurons from young SD rats and got abundant numbers and good status of EPCs and neurons.2.We found that constitutive addition of NT-3(200ng/ml)conditional medium could accelerate proliferative rates of EPCs in vitro.3.We found that constitutive addition of NT-3(200ng/ml)conditional medium could improve differentiate capacity of EPCs in vitro.Conclusions: We conclude that high dose and constitutive addition of NT-3 conditional medium could accelerate proliferative rates and differentiate capacity.Objective: Our research focuses on identifying the role of neurotrophins-3(NT-3)on proliferation,differentiation and function of bone marrow derived endothelial progenitor cells(BM-EPCs),we also check the effect of NT-3 treated endothelial progenitor cells(EPCs)on lower limb ischemia in rats.Methods: We first establish lower limb ischemia in rats,and then check the effect of transplantation of NT-3 treated EPCs on treating lower limb ischemia in rats.Results: 1.We established an animal disease model,which related to lower limb ischemia in rats.2.Through transplantation experiments,we found NT-3 conditional medium treated EPCs could increase blood flow in animal models.3.Through immunohistochemistry,we found that NT-3 conditional medium could improve differentiate capacity of EPCs in vivo.4.Through ELISA experiments,we found NT-3 conditional medium could promote EPCs to secrete VEGF165.5.Through immunostaining,we found NT-3 conditional medium could decrease percentage of apoptotic EPCs.Conclusions: We conclude that high dose and constitutive addition of NT-3 conditional medium could accelerate proliferative rates and differentiate capacity.NT-3 treated EPCs could be applied in treating low limb ischemia in rats,partially due to more secretion of VEGF165.