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- W3028990057 abstract "ObjectiveTo study the transduction efficiency of expressing human neprilysin by using a lentiviral vector (Lenti-NEP) in mouse embryonic neural stem cells (NSC) in vitro.MethodsPrimary NSC were harvested from C57BL/6J pregnant mouse at embryonic day 11.5 and transducted with Lenti-NEP.Immunofluorescent stainingand Western blot were performed to detect NEP protein expression in NSC.Degradation of amyloid beta 1-40 (Aβ1-40) by NEP protein transduced with Lenti-NEP in NSC was analyzed using ELISA and HPLC.ResultsOver 90% NSC were successfully transduced with Lenti-NEP via observation of fused protein green fluorescent protein under the microscopy.Expressions of NEP transduced with Lenti-NEP in NSC and of the markers of NSC (nestin) and neuron (MAP2).The enzyme activity of 2.5μg (21.00±2.51) and 1.0μg (15.00±0.54) NEP on degrading Aβ1-40 was shown to improve significantly compared to 0.5μg NEP (8.00±0.81，t=40.4 and 12.7，respectively，both P<0.01).The activity of NEP was inhibited in the presence of 50μmol/L phosphoramidon (0.5μg:0.08±0.01;1.0μg:0.04±0.01;2.5μg:0.05±0.01, t=17.2，51.3 and 14.1，respectively，all P<0.01).The hydrolytic cleavage on degrading Aβ1-40 by NEP was 11.4%，28.4% and 93.7% with incubation for 1 h，4 h and 12 h，respectively.ConclusionsLentiviral vector successfully delivers NEP gene to NSC in vitro.Targeting on NEP and NSC may provide potential therapeutic tool for Alzheimer’s disease.Key words: Neprilysin; Neural stem cells; Lentivirus; Transfection; Amyloid betaprotein" @default.
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- W3028990057 date "2013-01-08" @default.
- W3028990057 modified "2023-09-28" @default.
- W3028990057 title "The efficiency of expressing human neprilysin by using lentiviral vector transduction in neural stem cells" @default.
- W3028990057 doi "https://doi.org/10.3760/cma.j.issn.1006-7876.2013.01.006" @default.
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