Page 93 - 2020年20期

基于PI3K/Akt/GSK3β信号通路的芥子酸抗Aβ 1-42致PC12细胞损

伤的机制研究 Δ

1 #
3
2
1＊
薛 迪 ，刘宇超 ，贾永明 ，汪 娜 ，刘学伟 （1.齐齐哈尔医学院药学院，黑龙江 齐齐哈尔 161006；2.齐齐哈
1
尔医学院科研处，黑龙江 齐齐哈尔 161006；3.齐齐哈尔医学院基础医学院，黑龙江 齐齐哈尔 161006）
中图分类号 R285 文献标志码 A 文章编号 1001-0408（2020）20-2519-05
DOI 10.6039/j.issn.1001-0408.2020.20.16
摘 要 目的：基于磷脂酰肌醇-3-激酶（PI3K）/蛋白激酶B（Akt）/糖原合成激酶3β（GSK3β）信号通路探讨芥子酸（SA）抗β淀粉样
蛋白1-42（Aβ1-42 ）致PC12细胞损伤的作用机制。方法：将大鼠PC12细胞随机分为对照组、Aβ组（Aβ1-42 2 μmol/L）、Aβ+SA组（Aβ1-42
2 μmol/L +SA 100 μmol/L）、Aβ+SA+LY组[Aβ1-42 2 μmol/L +SA 100 μmol/L+LY294002（PI3K抑制剂）10 μmol/L]、Aβ+LY组（Aβ1-42 2
μmol/L+LY294002 10 μmol/L）、LY 组（LY294002 10 μmol/L）。除对照组、LY 组外，其余各组细胞均以 Aβ 1-42复制损伤模型。培养
24 h 后，使用显微镜观察各组细胞的形态，采用 MTT 法检测各组细胞的存活率；采用 Western blotting 法检测各组细胞 PI3K、
p-PI3K、Akt、p-Akt、GSK3β、p-GSK3β蛋白的表达情况。结果：与对照组比较，Aβ组细胞数量变少、部分突触断裂消失，其存活率以
及p-PI3K/PI3K、p-Akt/Akt、p-GSK3β/GSK3β比值均显著降低（P＜0.05或P＜0.01）。与Aβ 组比较，Aβ+SA组细胞变圆、突触变多，
其存活率以及p-PI3K/PI3K、p-Akt/Akt、p-GSK3β/GSK3β比值均显著升高（P＜0.05）。与Aβ+SA组比较，Aβ+SA+LY组细胞部分突
触断裂，其存活率以及p-PI3K/PI3K、p-Akt/Akt、p-GSK3β/GSK3β比值均显著降低（P＜0.05）；Aβ+LY组细胞碎片较多，其存活率虽
有下降但差异无统计学意义，且 p-PI3K/PI3K、p-Akt/Akt、p-GSK3β/GSK3β比值亦无明显变化（P＞0.05）。单独给予 LY294002 对
PC12 细胞的形态、存活率以及 p-PI3K/PI3K、p-Akt/Akt、p-GSK3β/GSK3β比值均无显著影响（P＞0.05）。结论：SA 可能通过激活
PI3K/Akt/GSK-3β信号通路对Aβ1-42诱导的PC12 细胞损伤发挥保护作用。
关键词 芥子酸；β淀粉样蛋白；磷脂酰肌醇-3-激酶/蛋白激酶B/糖原合成激酶3β信号通路；PC12细胞

Study on the Mechanism of Sinapic Acid against PC12 Cell Injury Induced by Aβ 1-42 Based on PI3K/Akt/
GSK3β Signaling Pathway
XUE Di ，LIU Yuchao ，JIA Yongming ，WANG Na ，LIU Xuewei（1. School of Pharmacy，Qiqihar Medical
1
1
1
2
3
University，Heilongjiang Qiqihar 161006，China; 2. Office of Academic Research，Qiqihar Medical University，
Heilongjiang Qiqihar 161006，China; 3. School of Basic Medicine，Qiqihar Medical University，Heilongjiang
Qiqihar 161006，China）
ABSTRACT OBJECTIVE：To investigate the mechanism of sinapic acid（SA）against PC12 cell injury induced by Amyloid β 1-42
protein（Aβ 1-42 ）based on PI3K/Akt/GSK3β signaling pathway. METHODS：PC12 cells of rats were randomly divided into control
group，Aβ group（Aβ 1-42 2 μmol/L），Aβ+SA group（Aβ 1-42 2 μmol/L+SA100 μmol/L），Aβ+SA+LY group [Aβ 1-42 2 μmol/L+SA 100
μmol/L+LY294002（PI3K inhibitor）10 μmol/L]，Aβ+LY group（Aβ 1-42 2 μmol/L+LY294002 10 μmol/L）and LY group（LY294002
10 μmol/L）. Except for control group and LY group，the cells of other groups were replicated the damage model with Aβ 1-42. After
24 hours of culture，the morphology of cells was obsened in each group with a microscope，and MTT assay was adopted to
determine the cell viability of PC12 cells in each group. Western blotting assay was used to detect the expression of PI3K，p-PI3K，
Akt，p-Akt，GSK3β and p-GSK3β in cells of each group. RESULTS：Compared with control group，the number of cells decreased
and some synaptic breaks disappeared in Aβ group while cell viability，ratio of p-PI3K/PI3K，p-Akt/Akt and p-GSK3β/GSK3β in
Aβ group were decreased significantly（P＜0.05 or P＜0.01）. Compared with Aβ group，the cells became round and synapses
became more in Aβ+SA group while cell viability，the ratio of p-PI3K/PI3K，p-Akt/Akt and p-GSK3β/GSK3β were increased
significantly（P＜0.05）. Compared with Aβ+SA group，some synaptic breaks occurred in Aβ+SA+LY group while cell viability，
the ratio of p-PI3K/PI3K，p-Akt/Akt and p-GSK3β/GSK3β were decreased significantly（P＜0.05）；Aβ+LY group had more cell
debris，and the cell viability was decreased，but the difference was not significant，and the ratio of p-PI3K/PI3K，p-Akt/Akt and
p-GSK3 β/GSK3 β had no significant change （P＞0.05）;
Δ 基金项目：黑龙江省省属高等学校基本科研业务费科研项目
LY294002 alone had no significant effect on morphology，cell
（No.2017-KYYWF-0698）
viability and the ratio of p-PI3K/PI3K，p-Akt/Akt or p-GSK3β/
＊研究实习员，硕士。研究方向：天然药物防治神经退行性疾病
GSK3 β （P＞0.05）. CONCLUSIONS： SA may play a
的 活 性 成 分 筛 选 及 作 用 机 制 。 电 话 ：0452-2663615。 E-mail：
282832817@qq.com protective role against PC12 cell injury induced by A β 1-42
# 副研究员，硕士生导师，博士。研究方向：天然药物防治神经退 through activating PI3K/Akt/GSK-3β.
行性疾病的活性成分筛选及作用机制。电话：0452-2663804。E-mail： KEYWORDS Sinapic acid；β-amyloid protein；PI3K/Akt/
lxw_qmu@126.com GSK3β signaling pathway；PC12 cell

中国药房 2020年第31卷第20期 China Pharmacy 2020 Vol. 31 No. 20 ·2519 ·