Aspirin eugenol ester ameliorates LPS-induced inflammatory responses in RAW264.7 cells and mice

Xu Liu; Qi Tao; Youming Shen; Xiwang Liu; Yajun Yang; Ning Ma; Jianyong Li

doi:10.3389/fphar.2023.1220780

Aspirin eugenol ester ameliorates LPS-induced inflammatory responses in RAW264.7 cells and mice

Front Pharmacol. 2023 Aug 29:14:1220780. doi: 10.3389/fphar.2023.1220780. eCollection 2023.

Authors

Xu Liu¹, Qi Tao², Youming Shen³, Xiwang Liu², Yajun Yang², Ning Ma¹, Jianyong Li²

Affiliations

¹ Hebei Veterinary Biotechnology Innovation Center, College of Veterinary Medicine, Hebei Agricultural University, Baoding, Hebei, China.
² Key Lab of New Animal Drug Project of Gansu Province, Key Lab of Veterinary Pharmaceutical Development of Ministry of Agriculture and Rural Affairs, Lanzhou Institute of Husbandry and Pharmaceutical Science of Chinese Academy of Agricultural Sciences, Lanzhou, Gansu, China.
³ Quality Inspection and Test Center for Fruit and Nursery Stocks, Ministry of Agriculture and Rural Affairs (Xingcheng), Research Institute of Pomology Chinese Academy of Agricultural Sciences, Xingcheng, Liaoning, China.

Abstract

Introduction: Inflammation is a defensive response of the body and the pathological basis of many diseases. However, excessive inflammation and chronic inflammation impair the homeostasis of the organism. Arachidonic acid (AA) has a close relationship with inflammation and is the main mediator of the pro-inflammatory response. Based on the prodrug principle, the new pharmaceutical compound aspirin eugenol ester (AEE) was designed and synthesized. However, the effects of AEE on key enzymes, metabolites and inflammatory signaling pathways in the AA metabolic network have not been reported. Methods: In this study, the anti-inflammation effects of AEE were first investigated in mice and RAW264.7 cells in LPS induced inflammation model. Then, the changes of the key enzymes and AA metabolites were explored by RT-PCR and targeted metabolomics. Moreover, the regulatory effects on NF-kB and MAPKS signaling pathways were explored by Western Blotting. Results: Results indicated that AEE significantly reduced the number of leukocyte and increased the lymphocyte percentage. AEE decreased the expression levels of IL-1β, IL-6, IL-8 and TNF-α both in vivo and in vitro. In the liver of mice, AEE downregulated the levels of AA, prostaglandin D₂ (PGD₂) and upregulated 12- hydroxyeicosatetraenoic acid (12-HETE). However, the changes of PGE₂, PGF_2α, 6-keto-prostaglandin F_1α (6-KETO-PGF_1α), 9-hydroxy-octadecenoic acid (9- HODE), 13-HODE, 15-HETE, docosahexaenoic acid (DHA) and thromboxane B₂ (TXB₂) were not significant. Additionally, it was found that AEE decreased the relative mRNA expression levels of p65 and p38 and the ratio of p-p65/p65. Discussion: It was concluded that AEE might inhibit the LPS-induced inflammatory response through the regulation of AA metabolism. This study provides the theoretical foundation for the development of AEE as a medicinal anti-inflammatory drug.

Keywords: Arachidonic acid; MAPK; NF-κB; UPLC-MS/MS; aspirin eugenol ester; inflammation.

Grants and funding

This work was supported by the Science–technology innovation engineering of CAAS (25-LZIHPS-02), Natural Science Foundation of Hebei Province (C2021204035).