Coumarin and eugenol ameliorate LPS-induced inflammation in RAW 264.7 cells via modulating the NLRP3 inflammasome pathway

Abstract

Objective: To investigate the underlying mechanism of anti-inflammatory action of coumarin and eugenol in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. Methods: RAW 264.7 cells were treated with 2.5 µg/mL of LPS, 50 µM of coumarin, and 50 µM eugenol for 24 h. The viability of the cells was assessed using MTT assay. The production of nitric oxide was determined using Griess reagent and DCFH-DA was used to measure the production of reactive oxygen species. The protein expression of NLRP3, IL-1β, NF-κB, and cyclooxygenase 2 was assessed using Western blot analysis. Results: Coumarin and eugenol showed anti-inflammatory effects against LPS-induced inflammatory response by ameliorating the expression of NLRP3 inflammasome and NF-κB, which further led to a subsequent reduction in IL-1β, nitric oxide, and reactive oxygen species. Conclusions: Coumarin and eugenol exert their anti-inflammatory activities by modulating the NLRP3 inflammasome pathway and NF-κB. These compounds may have promising therapeutic applications for the treatment of various inflammatory diseases.

Keywords

Inflammation / Cytokines / Coumarin / Eugenol / NLRP3 / LPS

Acknowledgments

The authors would like to thank Ms Gitika Sharma for her support during the study. JB thanks DRDO for financial support in the form of a Junior Research Fellowship.

Conflict of interest statement

The authors declare no competing interests.

Funding

This work was supported by the Defence Institute of Physiology and Allied Sciences.

Data availability statement

The data supporting the findings of this study are available from the corresponding authors upon request.

Authors’ contributions

KPM designed the study; JB performed the experiments and collected the data; JB and KPM analyzed the data; JB and KPM drafted the manuscript; KPM and SS provided the resources and materials; JB, SS and KPM edited the manuscript. All authors have read and agreed to the published version of the manuscript.

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