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OBJECTIVE TO investigate the effect of high dose β-amyloid 1-42 oligomers(AβO1-42) on adult hippocampal neural stem/progenitor cells(NSC) in vitro.METHODSNSC was treated with AβO1-42(3-10 μmol·L-1) for 5 d.Measurements of cell proliferation were performed using a bromodeoxyuridine(5-bromo-2-deoxyuridine,BrdU) incorporation assay.Intracellular reactive oxygen species(ROS) levels were monitored by using 2′,7′-dichlorofluorescin diacetate(DCFH-DA).NSC senescence was evaluated by using SA-β-gal(senescence-associated β-galactosidase) assay.Antioxidant N-acetyl-L-cysteine(NAC) and p38 inhibitor SB20358 were tested for their effects on the AβO1-42-induced NSC senescence.The expressions of p38 kinase phosphorylation and prosenescence molecules p16INK4a and Rb were determined by Western blotting.RESULTS High dose AβO1-42 dose-dependently inhibited NSC proliferation,and enhanced intracellular ROS levels,contributing to NSC senescence(P<0.05).In addition,AβO1-42 upregulated p38 kinase phosphorylation,p16INK4a and Rb protein expression.Either 1 mmol·L-1 NAC or 10 μmol·L-1 SB203580 attenuated the levels of AβO1-42-enhanced p16INK4a and Rb protein expression,and prevented AβO1-42-induced NSC senescence.CONCLUSION High dose AβO1-42 induce NSC senescence via ROS-p38-p16-Rb pathway,thereby inhibiting adult neurogenesis.
OBJECTIVE TO investigate the effect of high dose β-amyloid 1-42 oligomers (AβO 1-42) on adult hippocampal neural stem / progenitor cells (NSC) in vitro. METHODSNSC was treated with AβO 1-42 (3-10 μmol·L -1 ) for 5 d. Measurements of cell proliferation were performed using a bromodeoxyuridine (5-bromo-2-deoxyuridine, BrdU) incorporation assay.Intracellular reactive oxygen species (ROS) levels were monitored by using 2 ’, 7’-dichlorofluorescin diacetate -DA) .NSC senescence was evaluated by using SA-β-gal (senescence-associated β-galactosidase) assay. Antioxidant N-acetyl-L-cysteine (NAC) and p38 inhibitor SB20358 were tested for their effects on the AβO 1-42 -induced NSC senescence. These expressions of p38 kinase phosphorylation and prosenescence molecules p16INK4a and Rb were determined by Western blotting. RESULTS High dose AβO 1-42 dose-dependently inhibited NSC proliferation, and enhanced intracellular ROS levels, contributing to NSC senescence (P <0.05 In addition, AβO1-42 upregulated p38 kina se phosphorylation, p16INK4a and Rb protein expression. Either 1 mmol·L-1 NAC or 10 μmol·L-1 SB203580 attenuated the levels of AβO1-42-enhanced p16INK4a and Rb protein expression, and prevented AβO1-42-induced NSC senescence. CONCLUSION High dose AβO1-42 induce NSC senescence via ROS-p38-p16-Rb pathway, thereby inhibiting adult neurogenesis.