Exploring the In Vitro Anti-Inflammatory Effect and In Silico Toxicity Profile of Curcuma aeruginosa Roxb. Extract in RAW 264.7 Macrophages

Abstract

Chronic inflammation is implicated in numerous diseases, and targeting pro-inflammatory pathways is a promising therapeutic approach. Curcuma aeruginosa Roxb. known for its traditional medicinal uses, contains sesquiterpenes that may offer anti-inflammatory benefits. This study examines the anti-inflammatory properties of ethanol extract from C. aeruginosa Roxb. rhizome on nitric oxide (NO) production in LPS-stimulated RAW 264.7 macrophages using in vitro and in silico approaches. The in vitro analysis was conducted by treating RAW 264.7 cells with varying concentrations of the extract (15, 30, and 60 µg/mL), followed by NO measurement using the Griess assay to evaluate inhibition. The IC50 value of the extract was determined to be 29.03 ± 4.37 µg/mL. Notably, cell viability analysis using the WST-1 assay confirmed that these treatment concentrations did not induce toxicity in the cells. In the in silico analysis, 17 sesquiterpene compounds identified from the extract were screened for bioactivity and toxicity, then tested in molecular docking and molecular dynamics simulations with primary inflammatory proteins, including iNOS, IKKβ, ERK2, JNK1, and p38. Molecular docking and molecular dynamics results highlighted three key compounds, which were dehydrochromolaenin, pyrocurzerenone, and turmeronol B at exhibited strong binding affinities, particularly with iNOS, indicating stable interactions with significant anti-inflammatory potential. These findings suggest that C. aeruginosa rhizome extract effectively reduces NO production in vitro and demonstrates molecular interactions that inhibit inflammatory mediators in silico. This extract holds potential as a natural anti-inflammatory agent with a multifaceted mechanism of action against inflammatory signaling pathways.