Curcumin improves autophagy in N2a/APP695swe cells via elevating the expressions of Atg proteins

Kang Qingmei; Liang Jie; Zhang Xiong; Zhou Fanlin; Li Dan; Gao Minna; Li Yu

doi:10.13406/j.cnki.cyxb.001249

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Curcumin improves autophagy in N2a/APP695swe cells via elevating the expressions of Atg proteins

更新时间：2024-01-17

- Curcumin improves autophagy in N2a/APP695swe cells via elevating the expressions of Atg proteins
- Journal of Chongqing Medical University Vol. 42, Issue 5, Pages: 481-486(2017)
- 作者机构：
  
  1. 重庆医科大学基础医学院病理学教研室
  2. 重庆医科大学神经科学研究中心
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.001249
  CLC： R285
- Published：2017，
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Kang Qingmei, Liang Jie, Zhang Xiong, et al. Curcumin improves autophagy in N2a/APP695swe cells via elevating the expressions of Atg proteins. [J]. Journal of Chongqing Medical University 42(5):481-486(2017)
DOI：

Kang Qingmei, Liang Jie, Zhang Xiong, et al. Curcumin improves autophagy in N2a/APP695swe cells via elevating the expressions of Atg proteins. [J]. Journal of Chongqing Medical University 42(5):481-486(2017) DOI： 10.13406/j.cnki.cyxb.001249.

摘要

目的:全面研究姜黄素(curcumin)对N2a/APP695swe细胞自噬作用的影响

进一步探讨姜黄素调节N2a/APP695swe细胞自噬的可能机制。方法:将细胞分为4组:野生型对照组(WT组)、空白对照组(Control组)、溶剂对照组(Sham组)、姜黄素处理组(Curcumin组

5μmol/L)。Western blot分别检测自噬标志性蛋白LC3Ⅱ和底物蛋白SQSTM1的表达

以及Atg蛋白Beclin1、Atg5、Atg7、Atg12、Atg16L1的表达;免疫荧光法进一步验证SQSTM1蛋白的表达。各组分别转染细胞自噬体指示探针(GFPLC3)和溶酶体红色探针(Lyso Red)后

激光共聚焦显微镜观察自噬体和自噬溶酶体的数量检测自噬流。结果:Western blot结果表明

Curcumin组LC3Ⅱ蛋白表达量较对照组明显增高(0.59±0.03 vs.0.11±0.01

P=0.000);同时

Curcumin组SQSTM1蛋白表达量较对照组明显降低(0.19±0.04 vs.0.66±0.05

P=0.000)。免疫荧光实验再次验证了SQSTM1蛋白的这个表达趋势。共聚焦显微镜观察发现

与对照组相比

Curcumin组细胞内自噬体数量增加(154.67±7.37 vs.89.00±7.21

P=0.000)

同时自噬溶酶体数量也显著增加(84.67±12.50 vs.42.33±6.51

P=0.000)。Western blot进一步检测发现

与对照组相比

Curcumin组Atg蛋白Beclin1、Atg7、Atg16L1的表达均明显增加(0.55±0.09 vs.0.10±0.02

P=0.000;0.89±0.12 vs.0.39±0.07

P=0.000;0.36±0.07vs.0.13±0.02

P=0.003)

而Atg5和Atg12改变差异无统计学意义(0.79±0.12 vs.0.79±0.10

P=0.470;0.94±0.13 vs.0.87±0.16

P=0.587)。结论:姜黄素增强N2a/APP695swe细胞的自噬水平

其机制可能与提高自噬体形成相关蛋白Beclin1、Atg7、Atg16L1的表达有关。

Abstract

Objective:To explore the effects of curcumin on autophagy of N2a/APP695 swe cells and the underlying mechanisms.Methods:The N2a/WT and N2a/APP695 swe cells were involved

and were divided into four groups:wild type（WT）group

control group

sham group and curcumin group（5 μmol/L）. Western blot was carried out to detect the expressions of LC3Ⅱ

SQSTM1

and the autophagy-related genes（Atg）including Beclin1

Atg5

Atg7

Atg12

and Atg16L1 proteins. Immunofluorescence was used to test the expression of SQSTM1 protein

too. After transfection of p SELECT-GFP-LC3 and Lyso Red into the cells of four groups

respectively

confocal microscope was performed to observe the difference of the number of autophagosomes and autolysosomes in each group.Results:Western blot demonstrated that compared with those of control group

the expressions of LC3 Ⅱ protein were significantly increased in curcumin group（0.59±0.03 vs. 0.11±0.01

P=0.000）. But the expression of SQSTM1 protein was attenuated in curcumin group than that in control group

simultaneously（0.19±0.04 vs. 0.66±0.05

P=0.000）. And immunofluorescence also verified this trend of SQSTM1 protein. Then confocal microscope showed that the number of autophagosomes and the quantity of autolysosomes were improved in curcumin group

compared with those in control group（154.67±7.37 vs. 89.00±7.21

P=0.000;84.67±12.50 vs. 42.33±6.51

P =0.000）. At last

Western blot further found that the expressions of Beclin1

Atg7

and Atg16L1 at protein levels were increased in curcumin group when compared with those of control group（0.55±0.09 vs. 0.10±0.02

P=0.000;0.89±0.12 vs. 0.39±0.07

P=0.000;0.36±0.07 vs. 0.13±0.02

P=0.003）. However

there was no significant difference in the two groups at the levels of Atg5 and Atg12 proteins（0.79±0.12 vs. 0.79±0.10

P=0.470;0.94±0.13 vs. 0.87±0.16

P=0.587）. Conclusion:Curcumin plays a key role in elevating autophagy in N2a/APP695 swe cells and the potential mechanisms is associated with Atg proteins including Beclin1

Atg7

and Atg16L1.

关键词

姜黄素自噬阿尔茨海默病Beclin1Atg7Atg16L1

Keywords

curcuminautophagyAlzheimer’s diseaseBeclin1Atg7Atg16L1

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