上海交通大学学报（医学版）

›› 2013, Vol. 33 ›› Issue (4): 396-.doi: 10.3969/j.issn.1674-8115.2013.04.003

• 论著（基础研究） • 上一篇    下一篇

丁苯酞对β淀粉样蛋白诱导的U87细胞自噬性死亡的影响

王红梅, 张 婷, 李 强, 陈荣富, 孙晓江   

  1. 上海交通大学附属第六人民医院神经内科, 上海 200233
  • 出版日期:2013-04-28 发布日期:2013-05-03
  • 通讯作者: 孙晓江, 电子信箱: sunxj155@sohu.com。
  • 作者简介:王红梅（1975—）, 女, 主治医师, 博士； 电子信箱: dd-tt12345@163.com。
  • 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

WANG Hong-mei, ZHANG Ting, Li Qiang, CHEN Rong-fu, SUN Xiao-jiang   

  1. Department of Neurology, the Sixth People´s Hospital, Shanghai Jiaotong University, Shanghai 200233, China
  • Online:2013-04-28 Published:2013-05-03
  • Supported by:

    National Natural Science Foundation of China, 81100953, 31171014, 31100783

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摘要：

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。

关键词: 丁苯酞, 阿尔茨海默病, β淀粉样蛋白, 氧化应激, 自噬性死亡

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