ABSTRACT
Objective To explore the effect of the complex containing neuron stem cells(NSCs)and endothelial progenitor cells(EPCs)after being transplanted to ischemia and reperfusion rat model on the differentiation of neural stem cells to neuron and synapse formation.MethodsTotally 300 SD adult male rats were randomly divided into sham operation group,middle cerebral artery occlusion(MACO)model group,MACO+NSCs group,MACO+EPCs group,and MACO+complex group,and every group were further divided into 6 subgroups according to time points 1,3,7,14,30 and 60 d after operation,NSCs were isolated from the hippocampus of neonatal SD rats born in 24 h and identified with Nestin staining.EPCs were obtained from the artery blood of SD rats and verified with immunocytochemical stainings of CD31 and CD34 after culture.NSCs and EPCs were cultured together to produce the complex with aid of laminin.Then normal saline,the NSCs,EPCs,or complex(2?1010/L)were transplanted into the ischemia penumbra of corresponding model rat brain,sham control or MACO rats.The differentiation of NSCs,the expression of P38,synapsin-I,mRNA of connexin 43 were observed with double label immunofluorescence technique,immunohistochemical method,RT-PCR and Western blot analysis in ischemic penumbra.ResultsCompared with other groups,there were more double Brdu and neural special enolase positive cells in MACO+complex group with the increased expression of P38.RT-PCR and Western blot analysis indicated that MACO+complex group had significantly higher expressions of synapsin-I at mRNA and protein levels in ischemic penumbra,and so did the mRNA expression of connexin 43.ConclusionThe transplantation of the complex of NSCs and EPCs improves the NSCs differentiation to neuron,synapse formation and reconstruction of neural network.
ABSTRACT
Objective To explore the relationship between angiogenesis and ischemic neurological deficit repair through exogenous interventions,and investigate optimal intervention routes for angiogenesis in cerebral ischemia rats.Methods Totally 240 SD rats were randomly divided into sham operation group,ischemic control group,ischemia and vascular endothelial growth factor(VEGF) group [1 653.3 ng/(g?d),i.p.,for 10 d],ischemia and endothelial progenitor cell transplantation(EPCs) group(2?1010/L,10 ?l),ischemia and anti-VEGF monoclonal antibody group [3 mg/(kg?d),i.p.,for 10 d].Cerebral microvessel density,somatosensory evoked potentials(SEP) and motor evoked potentials(MEP) and behavior were observed in 1,3,7,14,30 and 60 d after injury.Results New blood vessel density of ischemia and VEGF group,ischemia and EPCs group were significantly higher than ischemic control group in all time points besides 1 d after injury(P
ABSTRACT
Objective To investigate the effect of endothelial progenitor cells (EPCs) which complexed with neuron stem cells (NSCs) after transplanted into ischemia and reperfusion (I/R) rat model on the proliferation and apoptosis of the NSCs and vasal construction. Methods Totally 150 SD adult male rats were randomly and equally divided into 5 groups, sham-operation group, I/R group, I/R+NSCs group, I/R+EPCs group, and I/R+NSCs+EPCs group. The rats were further divided into 5 subgroups according to the time points of 3, 7, 14, 30 and 60 d after transplantation. I/R rat model was established by reversiblyligating middle cerebral artery occlusion. NSCs were derived from the hippocampus of SD rats born in 24 h and identified with Nestin staining. EPCs were obtained from the artery blood of SD rats and verified with immunocytochemical stainings of CD31, CD34 and KDR after culture. The complex of NSCs and EPCs was produced with aid of laminin. Then the complex were transplanted into the ischemia penumbra of corresponding model rat brain. The proliferation and apoptosis of NSCs, and the fomation of new vessels were observed through immunohistochemistry and double-labeled immunofluorescence staining. Results The proliferation of NSCs was increased, apoptosis cells were decreased and the new vessels were raised in the complex transplantation when compared with single NSCs or EPCs transplantation groups. Conclusion The complex of NSCs and EPCs transplantation improves the proliferation of NSCs, suppresses cell apoptosis and promotes the formation of new vessels.