丁苯酞对β淀粉样蛋白诱导的U87细胞自噬性死亡的影响
网络出版日期: 2013-05-03
基金资助
国家自然科学基金(81100953, 31171014, 31100783)
Effect of dl-3-n-butylphthalide on autophagic cell death in Aβ-treated U87 cells
Online published: 2013-05-03
Supported by
National Natural Science Foundation of China, 81100953, 31171014, 31100783
目的 观察丁苯酞对β淀粉样蛋白(Aβ1-42)处理后U87细胞自噬性死亡的影响。方法 方法将U87细胞分为Aβ组、Aβ+丁苯酞组和对照组(空白对照)。Aβ组细胞用Aβ1-42(20 μmol/L)处理24 h;Aβ+丁苯酞组细胞用丁苯酞(10 μmol/L)预处理0.5 h后再加入Aβ1-42(20 μmol/L)处理24 h。采用 MTT法测定细胞活力,检测Caspase 3活性,采用CM-H2 DCFDA检测胞内活性氧(ROS)水平,Western blotting检测LC3-Ⅰ和LC3-Ⅱ蛋白表达。结果 与对照组比较,Aβ组U87细胞活力显著下降(P<0.01);Aβ+丁苯酞组U87细胞活力显著高于Aβ组(P<0.05)。对照组、Aβ组和Aβ+丁苯酞组之间Caspase 3活性的差异均无统计学意义(P>0.05)。与对照组比较,Aβ组U87细胞ROS水平显著升高(P<0.01);Aβ+丁苯酞组U87细胞ROS水平显著低于Aβ组(P<0.01)。与对照组比较,Aβ组U87细胞LC3-Ⅱ/LC3-Ⅰ比值显著升高(P<0.01);Aβ+丁苯酞组U87细胞LC3-Ⅱ/LC3-Ⅰ比值显著低于Aβ组(P<0.01)。结论 丁苯酞通过抑制ROS介导的自噬性死亡发挥神经保护作用。
王红梅, 张 婷, 李 强, 等 . 丁苯酞对β淀粉样蛋白诱导的U87细胞自噬性死亡的影响[J]. 上海交通大学学报(医学版), 2013 , 33(4) : 396 . DOI: 10.3969/j.issn.1674-8115.2013.04.003
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.
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