木犀草素通过抑制Drp1活性改善自噬流过程减少脑缺血再灌注后氧化应激损伤

曾雪; 王龙; 赖玉洁; 陈波; 蒋航; 段晨阳; 张云东

doi:10.16016/j.1000-5404.202004273

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木犀草素通过抑制Drp1活性改善自噬流过程减少脑缺血再灌注后氧化应激损伤

神经医学 | 更新时间：2026-01-09

- 木犀草素通过抑制Drp1活性改善自噬流过程减少脑缺血再灌注后氧化应激损伤
- Luteolin attenuates oxidative stress in mice with cerebral ischemia-reperfusion by inhibiting Drp1 activation and promoting autophagy flux
- 第三军医大学学报 2020年42卷第18期页码：1777-1786
- 作者机构：
  
  1.401120 重庆，重庆医科大学附属第三医院神经疾病中心
  2.400038 重庆，陆军军医大学(第三军医大学)第一附属医院神经外科
  3.400042 重庆，陆军军医大学(第三军医大学)野战外科研究所战伤休克与输血研究室，创伤、烧伤与复合伤国家重点实验室
- 作者简介：
  
  [ 张云东，E-mail:zhangyundong1226@163.com ZHANG Yundong, E-mail:zhangyundong1226@163.com
- 基金信息：
  
  国家自然科学基金青年科学基金(81700429);国家自然科学基金青年科学基金(81801071)
- DOI：10.16016/j.1000-5404.202004273
  中图分类号： R282.71; R285.5; R743.31
- 收稿：2020-04-25，
  
  修回：2020-06-28，
  
  纸质出版：2020-09-30
- 稿件说明：
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曾雪, 王龙, 赖玉洁, 等. 木犀草素通过抑制Drp1活性改善自噬流过程减少脑缺血再灌注后氧化应激损伤[J]. 第三军医大学学报, 2020,42(18):1777-1786.

Xue ZENG, Long WANG, Yujie LAI, et al. Luteolin attenuates oxidative stress in mice with cerebral ischemia-reperfusion by inhibiting Drp1 activation and promoting autophagy flux[J]. Journal of Third Military Medical University, 2020, 42(18): 1777-1786.
曾雪, 王龙, 赖玉洁, 等. 木犀草素通过抑制Drp1活性改善自噬流过程减少脑缺血再灌注后氧化应激损伤[J]. 第三军医大学学报, 2020,42(18):1777-1786. DOI： 10.16016/j.1000-5404.202004273.

Xue ZENG, Long WANG, Yujie LAI, et al. Luteolin attenuates oxidative stress in mice with cerebral ischemia-reperfusion by inhibiting Drp1 activation and promoting autophagy flux[J]. Journal of Third Military Medical University, 2020, 42(18): 1777-1786. DOI： 10.16016/j.1000-5404.202004273.

摘要

目的

探究木犀草素(luteolin)对脑缺血再灌注氧化应激损伤的保护机制。

方法

通过靶向药物评分方法筛选出对脑缺血再灌注损伤具有潜在治疗效果的抗氧化剂并在大脑中动脉栓塞(middle cerebral artery occlusion，MCAO)小鼠模型中验证木犀草素的保护效果；在缺氧缺糖/复氧复糖(oxygen/glucose deprivation and re-oxygen/glucose，OGD/R)诱导的神经母细胞瘤SH-SY5Y细胞中使用不同浓度(10和50 μmol/L)木犀草素药物处理，检测MTT细胞活力及氧自由基ROS含量变化；通过免疫荧光、蛋白印迹等方法检测木犀草素处理后OGD/R诱导的SH-SY5Y细胞自噬流情况及线粒体形态变化，观察木犀草素对OGD/R后ROS生成和清除过程的影响。

结果

木犀草素对脑缺血再灌注靶向药物评分为96.86分。TTC染色结果显示木犀草素明显减少MCAO小鼠脑梗死面积(

0.05)。OGD/R后SH-SY5Y细胞MTT值约为正常对照组的50%，使用50 μmol/L的木犀草素处理后MTT值增加至83%(

0.05)，并且木犀草素显著减弱OGD/R诱导的ROS的产生，且呈剂量依赖性(

0.05)。激光共聚焦结果显示50 μmol/L木犀草素处理OGD/R诱导的SH-SY5Y细胞后自噬小体荧光强度增高64.6%(

0.05)，自噬小体与溶酶体共定位情况提高1.56倍(

0.05)。Western blot结果显示木犀草素明显抑制OGD/R后Drp1 Ser616磷酸化水平及Drp1线粒体转位(

0.05)，改善OGD/R后SH-SY5Y细胞线粒体形态(

0.05)。

结论

木犀草素对于脑缺血再灌注后氧化应激损伤具有明显保护作用，一方面它可以通过改善自噬流过程促进线粒体自噬小体的降解，加速ROS的清除效率；另一方面它还可以通过抑制Drp1活性保护线粒体形态，减少ROS的产生。

Abstract

Objective

To explore the mechanism by which luteolin alleviates oxidative stress in mice with cerebral ischemia-reperfusion.

Methods

We screened the antioxidants with potential therapeutic effect on cerebral ischemia-reperfusion injury using CMap dataset based on their targeted drug scores and verified the protective effect of luteolin in a mouse model of middle cerebral artery occlusion (MCAO). In a SH-SY5Y cell model of oxygen/glucose deprivation and reoxygenation (OGD/R)

we examined the effects of different concentrations of luteolin (10 and 50 μmol/L) on intracellular reactive oxygen species (ROS) content and cell viability using MTT assay. Immunofluorescence assay and Western blotting were performed to detect the changes in autophagy flux and mitochondrial morphology in response to luteolin treatment to assess the capacity of the cells for ROS production and clearance.

Results

The high targeted drug score (96.86) of luteolin suggested its potential protective effect against cerebral ischemia-reperfusion injury. TTC staining showed that treatment with luteolin significantly reduced cerebral infarction area in MCAO mice (

0.05). The MTT value of OGD/R-induced SH-SY5Y cells was about 50% of that of control cells

and was significantly increased to 83% after treatment with 50 μmol/L luteolin (

0.05). Luteolin also dose-dependently reduced ROS production in SH-SY5Y cells with OGD/R (

0.05). Confocal laser scanning microscopy showed that the fluorescence intensity of the autophagosomes was increased by 64.6% (

0.05) and the co-localization of autophagosomes and lysosomes increased by 1.56 folds after treatment with 50 μmol/L luteolin (

0.05). Western blotting demonstrated that luteolin significantly inhibited the phosphorylation of Drp1 at Ser616 and its mitochondrial translocation (

0.05)

and thereby improved the morphological changes of the mitochondria in OGD/R-induced SH-SY5Y cells (

0.05).

Conclusion

Luteolin has obvious protective effect against oxidative stress in mice with cerebral ischemia-reperfusion by promoting the degradation of mitophagosomes and autophagy flux to accelerate ROS clearance and by inhibiting the activity of Drp1 to protect mitochondrial morphology and reduce ROS production.

关键词

Keywords

references

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