Abstract:

Objective To observe the effect of dl-3-n-butylphthalide on the autophagic cell death in β-amyloid (Aβ1-42)-treated U87 cells. Methods U87 cells were divided into Aβ group, Aβ+dl-3-n-butylphthalide group and control group (blank control). Cells in Aβ group were treated with Aβ1-42 (20 μmol/L) for 24 h, and cells in Aβ+ dl-3-n-butylphthalide group were pre-incubated with dl-3-n-butylphthalide (10 μmol/L) for 0.5 h prior to treatment with 20 μmol/L Aβ1-42 for 24 h. Cell viability was detected by MTT, Caspase-3 activity of was examined, intracellular reactive oxygen species (ROS) was determined by CM-H2 DCFDA, and the expression of LC3-Ⅰ and LC3-Ⅱ protein was detected by Western blotting. Results The viability of U87 cells in Aβ group was significantly lower than that in control group (P<0.01), and the viability of U87 cells in Aβ+ dl-3-n-butylphthalide group was significantly higher than that in Aβ group (P<0.05). There was no significant difference in the Caspase 3 activity among Aβ group, Aβ+ dl-3-n-butylphthalide group and control group (P>0.05). The ROS level in U87 cells in Aβ group was significantly higher than that in control group (P<0.01), and the ROS level in U87 cells in Aβ+ dl-3-n-butylphthalide group was significantly lower than that in Aβ group (P<0.01). The LC3-Ⅱ/LC3-Ⅰ ratio in U87 cells in Aβ group was significantly higher than that in control group (P<0.01), and the LC3-Ⅱ/LC3-Ⅰ ratio in U87 cells in Aβ+ dl-3-n-butylphthalide group was significantly lower than that in Aβ group (P<0.01). Conclusion dl-3-n-butylphthalide exhibits neuroprotective effect through inhibition of ROS-mediated autophagic cell death in Aβ-treated U87 cells.

Key words: dl-3-n-butylphthalide, Alzheimer´s disease, β-amyloid, oxidative stress, autophagic cell death