Neuroprotective Effects of Eugenol Acetate Against Ischemic Stroke

Liqiu Chen,1,2,* Ran Zhang,1,2,* Jing Xiao,1,2,* Ying Liang,1,2 Zhen Lan,3 Yingao Fan,2 Xi Yu,2 Shengnan Xia,2– 5 Haiyan Yang,2– 5 Xinyu Bao,2– 5 Hailan Meng,1– 6 Yun Xu,1– 6 Linjie Yu,1– 6 Xiaolei Zhu1– 6 1Department of Neurology, Nanjing Drum Tower Hospital, Clinical Colleg...

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Main Authors: Chen L, Zhang R, Xiao J, Liang Y, Lan Z, Fan Y, Yu X, Xia S, Yang H, Bao X, Meng H, Xu Y, Yu L, Zhu X
Format: Article
Language:English
Published: Dove Medical Press 2025-01-01
Series:Journal of Inflammation Research
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Online Access:https://www.dovepress.com/neuroprotective-effects-of-eugenol-acetate-against-ischemic-stroke-peer-reviewed-fulltext-article-JIR
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author Chen L
Zhang R
Xiao J
Liang Y
Lan Z
Fan Y
Yu X
Xia S
Yang H
Bao X
Meng H
Xu Y
Yu L
Zhu X
author_facet Chen L
Zhang R
Xiao J
Liang Y
Lan Z
Fan Y
Yu X
Xia S
Yang H
Bao X
Meng H
Xu Y
Yu L
Zhu X
author_sort Chen L
collection DOAJ
description Liqiu Chen,1,2,* Ran Zhang,1,2,* Jing Xiao,1,2,* Ying Liang,1,2 Zhen Lan,3 Yingao Fan,2 Xi Yu,2 Shengnan Xia,2– 5 Haiyan Yang,2– 5 Xinyu Bao,2– 5 Hailan Meng,1– 6 Yun Xu,1– 6 Linjie Yu,1– 6 Xiaolei Zhu1– 6 1Department of Neurology, Nanjing Drum Tower Hospital, Clinical College of Nanjing University of Chinese Medicine, Nanjing, 210008, People’s Republic of China; 2Department of Neurology, Nanjing Drum Tower Hospital, Affiliated Hospital of Medical School, Nanjing University, Nanjing, 210008, People’s Republic of China; 3Department of Neurology, Nanjing Drum Tower Hospital, Clinical College of Nanjing Medical University, Nanjing, 210008, People’s Republic of China; 4State Key Laboratory of Pharmaceutical Biotechnology and Institute of Translational Medicine for Brain Critical Diseases, Nanjing University, Nanjing, 210008, People’s Republic of China; 5Jiangsu Provincial Key Discipline of Neurology, Nanjing, 210008, People’s Republic of China; 6Nanjing Neurology Medical Center, Nanjing, 210008, People’s Republic of China*These authors contributed equally to this workCorrespondence: Linjie Yu; Xiaolei Zhu, Department of Neurology, Nanjing Drum Tower Hospital, Clinical College of Nanjing University of Chinese Medicine, Nanjing, 210008, People’s Republic of China, Email yulinjie126@126.com; zhuquelee@126.comObjective: To explore the neuroprotective effect of Eugenol Acetate (EA) on post-stroke neuroinflammation and investigate the underlying mechanisms.Methods: For in vitro experiments, primary microglia were pre-incubated with EA for 2 hours, followed by lipopolysaccharide (LPS) stimulation for 24 hours or Oxygen-Glucose Deprivation (OGD) treatment for 4 hours. Real-time quantitative PCR, enzyme-linked immunosorbent assay (ELISA) and Western blot were performed to examine the expression levels of inflammatory cytokines in primary microglia. The activation of NF-κB signaling pathway was evaluated by immunofluorescence staining and Western blot. For in vivo experiments, middle cerebral artery occlusion (MCAO) was constructed to mimic ischemic brain injury on 8-week-old male C57BL/6J mice. The mice were continuously injected intraperitoneally with EA or vehicle after MCAO. Neurobehavioral tests and TTC staining were conducted to estimate the neurological deficits and infarct area. Moreover, the white matter integrity after MCAO was observed via immunofluorescence staining.Results: EA significantly reduced the expression of pro-inflammatory cytokines in LPS or OGD treated primary microglia, and inhibited LPS-induced activation of the NF-κB signaling pathway. In addition, EA alleviated ischemic brain injury and improved neuromotor function of MCAO mice. Furthermore, long-term neurological deficits and white matter integrity were improved by EA treatment after MCAO.Conclusion: EA alleviated ischemic injury and restored white matter integrity in MCAO mice, which might be associated with the inhibition of NF-κB signaling pathway in microglia. Therefore, EA might be a promising candidate for the treatment of ischemic stroke.Keywords: Eugenol Acetate, Ischemic Stroke, Neuroinflammation, NF-κB Signaling Pathway
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publishDate 2025-01-01
publisher Dove Medical Press
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series Journal of Inflammation Research
spelling doaj-art-cbd81f85bd3d403ea909ef17c34367482025-01-07T16:42:40ZengDove Medical PressJournal of Inflammation Research1178-70312025-01-01Volume 1813314698967Neuroprotective Effects of Eugenol Acetate Against Ischemic StrokeChen LZhang RXiao JLiang YLan ZFan YYu XXia SYang HBao XMeng HXu YYu LZhu XLiqiu Chen,1,2,* Ran Zhang,1,2,* Jing Xiao,1,2,* Ying Liang,1,2 Zhen Lan,3 Yingao Fan,2 Xi Yu,2 Shengnan Xia,2– 5 Haiyan Yang,2– 5 Xinyu Bao,2– 5 Hailan Meng,1– 6 Yun Xu,1– 6 Linjie Yu,1– 6 Xiaolei Zhu1– 6 1Department of Neurology, Nanjing Drum Tower Hospital, Clinical College of Nanjing University of Chinese Medicine, Nanjing, 210008, People’s Republic of China; 2Department of Neurology, Nanjing Drum Tower Hospital, Affiliated Hospital of Medical School, Nanjing University, Nanjing, 210008, People’s Republic of China; 3Department of Neurology, Nanjing Drum Tower Hospital, Clinical College of Nanjing Medical University, Nanjing, 210008, People’s Republic of China; 4State Key Laboratory of Pharmaceutical Biotechnology and Institute of Translational Medicine for Brain Critical Diseases, Nanjing University, Nanjing, 210008, People’s Republic of China; 5Jiangsu Provincial Key Discipline of Neurology, Nanjing, 210008, People’s Republic of China; 6Nanjing Neurology Medical Center, Nanjing, 210008, People’s Republic of China*These authors contributed equally to this workCorrespondence: Linjie Yu; Xiaolei Zhu, Department of Neurology, Nanjing Drum Tower Hospital, Clinical College of Nanjing University of Chinese Medicine, Nanjing, 210008, People’s Republic of China, Email yulinjie126@126.com; zhuquelee@126.comObjective: To explore the neuroprotective effect of Eugenol Acetate (EA) on post-stroke neuroinflammation and investigate the underlying mechanisms.Methods: For in vitro experiments, primary microglia were pre-incubated with EA for 2 hours, followed by lipopolysaccharide (LPS) stimulation for 24 hours or Oxygen-Glucose Deprivation (OGD) treatment for 4 hours. Real-time quantitative PCR, enzyme-linked immunosorbent assay (ELISA) and Western blot were performed to examine the expression levels of inflammatory cytokines in primary microglia. The activation of NF-κB signaling pathway was evaluated by immunofluorescence staining and Western blot. For in vivo experiments, middle cerebral artery occlusion (MCAO) was constructed to mimic ischemic brain injury on 8-week-old male C57BL/6J mice. The mice were continuously injected intraperitoneally with EA or vehicle after MCAO. Neurobehavioral tests and TTC staining were conducted to estimate the neurological deficits and infarct area. Moreover, the white matter integrity after MCAO was observed via immunofluorescence staining.Results: EA significantly reduced the expression of pro-inflammatory cytokines in LPS or OGD treated primary microglia, and inhibited LPS-induced activation of the NF-κB signaling pathway. In addition, EA alleviated ischemic brain injury and improved neuromotor function of MCAO mice. Furthermore, long-term neurological deficits and white matter integrity were improved by EA treatment after MCAO.Conclusion: EA alleviated ischemic injury and restored white matter integrity in MCAO mice, which might be associated with the inhibition of NF-κB signaling pathway in microglia. Therefore, EA might be a promising candidate for the treatment of ischemic stroke.Keywords: Eugenol Acetate, Ischemic Stroke, Neuroinflammation, NF-κB Signaling Pathwayhttps://www.dovepress.com/neuroprotective-effects-of-eugenol-acetate-against-ischemic-stroke-peer-reviewed-fulltext-article-JIReugenol acetateischemic strokeneuroinflammationnf-κb signaling pathway
spellingShingle Chen L
Zhang R
Xiao J
Liang Y
Lan Z
Fan Y
Yu X
Xia S
Yang H
Bao X
Meng H
Xu Y
Yu L
Zhu X
Neuroprotective Effects of Eugenol Acetate Against Ischemic Stroke
Journal of Inflammation Research
eugenol acetate
ischemic stroke
neuroinflammation
nf-κb signaling pathway
title Neuroprotective Effects of Eugenol Acetate Against Ischemic Stroke
title_full Neuroprotective Effects of Eugenol Acetate Against Ischemic Stroke
title_fullStr Neuroprotective Effects of Eugenol Acetate Against Ischemic Stroke
title_full_unstemmed Neuroprotective Effects of Eugenol Acetate Against Ischemic Stroke
title_short Neuroprotective Effects of Eugenol Acetate Against Ischemic Stroke
title_sort neuroprotective effects of eugenol acetate against ischemic stroke
topic eugenol acetate
ischemic stroke
neuroinflammation
nf-κb signaling pathway
url https://www.dovepress.com/neuroprotective-effects-of-eugenol-acetate-against-ischemic-stroke-peer-reviewed-fulltext-article-JIR
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