Objective:To construct a recombinant lentiviral vector that co-express ZsGreen and BDNF and establish an Adipose derived stem cell (ADSC) line with stable BDNF expression down- regulation.Methods: BDNF were designed and inserted into the lentiviral vector p HBLV-CMVIE-Zs Green-Puro. After identification by DNA sequencing, the lentiviral vectors carrying BDNF were packaged in 293 cells. The transfection efficiency was observed under fluorescence microscope;the lentiviral particles were collected to infect mouse ADSCs, and BDNF level were assessed using real-time PCR and Western blotting.Results: DNA sequencing demonstrated successful construction of the BDNF lentivirus vectors. Real- time PCR and Western blotting showed a high level BDNF mRNA and protein.Conclusion: We have successfully established an ADSC model with stable BDNF expression which establish the basis for the potential application of ADSCs-BDNF in the treatment of AD.
Alzheimer's disease(AD)has an unknown pathogenesis,which is one of the key factors leading to the death of the elderly.Recent studies have shown that glial cells,including microgli、astrocytes、oligo-dendrocytes and oligodendrocyte progenitor cells(NG2 glial cells),are closely related to the pathogenesis of AD.It is believed that activated microglial cells are very easy to induce neuritis and then lead to neurodegeneration.In this paper,the characteristics of the above four glial cells,their functions and their correlation with AD were reviewed,in order to provide a reference for the further study of the pathogenesis of AD.
Objective:To investigate whether Hainan papayas has protective effects in an Aimary neuron injury model and elucidate the underlying molecular mecha β 40-induced prnism.Methods:Cultured primary neurons from the dorsal root ganglia(DRG) of Sprague-Dawley(SD) rats were treated with 20 μM A culture medium β 40 peptide,100 μg/L Hainan papaya water extract,peptide plus extract,or for 24 h.Cell viability was measured by MTT assay,and neuronal apoptosis was evaluated by DAPI staining.ERK signaling pathway-associated molecule activation and changes in Bax expression were analyzed by Western blotting and immunofluorescence.Results:A cell viability rate of(44.11±6.59)% in the Ading different concentrations of the extract β 40 group was rescued to(79.13±6.64)% by ad.DAPI showed pyknotic nuclei in 39.5% of A β 40-treated cells;the fraction dropped to 17.4% in the 100 μg/L extract group.ERK phosphorylation was observed in the Aent with 100 μg/L extract.Hainan papaya water ex β 40 group but was ameliorated by pretreatmtract also prevented Ation of MEK,RSK1 and CREB associated with ERK signaling and d β 40-induced phosphorylaownregulated Bax expression in the neurons.Conclusion:The resultss suggest that Hainan papaya water extract has protective effects on neurons;the mechanism may be related to suppression of ERK signaling activation.
