Ethanol extracts of Cinnamomum migao H.W. Li attenuates neuroinflammation in cerebral ischemia-reperfusion injury via regulating TLR4-PI3K-Akt-NF-κB pathways

Wenze Wu; Libin Xu; Danyang Mu; Dequan Wang; Shaowen Tan; Linge Liu; Yubo Li; Huifang Chai; Yue Hou

doi:10.1016/j.jep.2024.119150

Ethanol extracts of Cinnamomum migao H.W. Li attenuates neuroinflammation in cerebral ischemia-reperfusion injury via regulating TLR4-PI3K-Akt-NF-κB pathways

J Ethnopharmacol. 2024 Nov 21:119150. doi: 10.1016/j.jep.2024.119150. Online ahead of print.

Authors

Wenze Wu¹, Libin Xu¹, Danyang Mu¹, Dequan Wang¹, Shaowen Tan¹, Linge Liu¹, Yubo Li², Huifang Chai³, Yue Hou⁴

Affiliations

¹ Key Laboratory of Bioresource Research and Development of Liaoning Province, College of Life and Health Sciences, National Frontiers Science Center for Industrial Intelligence and Systems Optimization, Key Laboratory of Data Analytics and Optimization for Smart Industry, Ministry of Education, Northeastern University, Shenyang 110169, China.
² School of Chinese Materia Medica, Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China. Electronic address: yaowufenxi001@sina.com.
³ School of Pharmacy, Guizhou University of Traditional Chinese Medicine, Guiyang, China. Electronic address: hfchai@126.com.
⁴ Key Laboratory of Bioresource Research and Development of Liaoning Province, College of Life and Health Sciences, National Frontiers Science Center for Industrial Intelligence and Systems Optimization, Key Laboratory of Data Analytics and Optimization for Smart Industry, Ministry of Education, Northeastern University, Shenyang 110169, China. Electronic address: houyue@mail.neu.edu.cn.

PMID: 39580135
DOI: 10.1016/j.jep.2024.119150

Abstract

Ethnopharmacological relevance: Cinnamomum migao H.W. Li, commonly known as migao (MG), is used in the Miao region of China for treating cardiovascular and cerebrovascular diseases, attributed to its detoxifying (Jiedu in Chinese), blood-activating (Huoxue in Chinese), and Qi-promoting (Tongqi in Chinese) effects. However, the therapeutic potential of MG for ischemic stroke (IS) has yet to be explored. Therefore, this study was to explore the protective effect of MG against cerebral ischemia-reperfusion injury caused by IS.

Aim of the study: The aim of this study was to investigate whether ethanol extract of MG (EEMG) attenuates cerebral ischemia-reperfusion injury, and explored the underlying mechanisms.

Materials and methods: Middle cerebral artery occlusion and reperfusion (MCAO/R) was established, and the efficacy of EEMG was evaluated using triphenyltetrazolium chloride (TTC), immunofluorescence, hematoxylin-eosin staining (HE) staining, and real-time quantitative PCR (qRT-PCR). Qualitative analysis of EEMG was analyzed for chemical composition by liquid chromatography-mass spectrometry (LC-MS). The molecular mechanism of EEMG was explored by metabolomics, network pharmacology, immunoblotting, immunofluorescence staining, gene knockdown, and agonist treatment.

Results: The results showed that EEMG can alleviate ischemic injury in MCAO/R-operated rats and neuronal damage of OGD/R-treated SH-SY5Y cells. Specifically, EEHGT inhibited the release of inflammatory factors and reversed serum metabolic profile disorders of MCAO/R rats. Network pharmacology analysis showed that the PI3K-Akt and NF-κB signaling pathways maybe involved in EEMG-mediated neuroprotective effects on ischemic injury and inhibition of inflammatory response. As we expected, EEMG can activate PI3K-AKT and suppress NF-kB signaling pathways both in MCAO/R-operated rats and OGD/R-treated BV2 cells. The results showed that knockdown of TLR4 abolished the EEMG-mediated inhibition on neuroinflammation in OGD/R-treated BV2 cells. After treating BV2 cells with the TLR4 agonist neoseptin 3, EEMG showed a trend toward inhibiting neuroinflammation but with no significant difference. Additionally, EEMG was found to improve liver injury caused by cerebral ischemia-reperfusion and associated with NF-κB signaling pathway in this study.

Conclusions: Collectively, this study demonstrated that EEMG attenuates neuroinflammation in cerebral ischemia-reperfusion injury via regulating TLR4-PI3K-Akt-NF-κB pathways.

Keywords: NF-κB; PI3K; TLR4; cinnamomum migao H.W. Li; ischemic stroke.