Investigating the anti-inflammatory potential of Eleusine coracana (L.) Gaertn. Seeds on LPS-induced RAW264.7 cell line with in-silico analysis

The purpose of this study is to evaluate the anti-inflammatory potential of the methanolic extract of Eleusine coracana seeds in RAW264.7 cells with in-silico analysis. The Phytoconstituents present in E. coracana seed extract were identified using GC–MS/MS and LC-MS/MS, and the results showed the presence of various anti-inflammatory compounds, including flavonoids, phenols, terpenoids, alkaloids, nucleosides, fatty acids, and their esters. Furthermore, LPS stimulation in RAW264.7 cells increased the ROS production from 23.36 ± 0.22 (control) to 43.35 ± 0.54, while E. coracana seed extract significantly reduced ROS to 31.23 ± 0.66 and 30.56 ± 1.35 at 10 and 50 μL/mL concentrations, respectively. Similarly, secretory levels of IL-6, IL-8, and TNF-α increased to 52.14 ± 6.06, 90.38 ± 0.54, and 2739.55 ± 3.85 pg/mL, respectively, in LPS-treated cells. However, treatment with E. coracana seed extract reduced these cytokines to 38.57 ± 5.05, 79.48 ± 0.72, and 2512.73 ± 7.07 pg/mL at 10 μL/mL, and further to 28.57 ± 1.01, 74.35 ± 0.72, and 2473.64 ± 28.92 pg/mL at 50 μL/mL. NF-κB expression was also suppressed, decreasing from 1.66 ± 0.10 in LPS-treated cells to 1.55 ± 0.14 at a 10 μL/mL concentration of E. coracana seed extract. Additionally, in-silico analysis showed that apiin exhibited the most favorable binding affinity (−7.2 kcal/mol) toward the key inflammatory target (TNF-α) when compared with other phytochemicals. Molecular dynamics simulations over 100 ns further confirmed the stability of the apiin-TNF-α complex, with both protein and ligand remaining stable throughout the simulation. Overall, this study highlights the first evidence from a cell line model and in silico methods that the methanolic extract of E. coracana seeds exhibits significant anti-inflammatory properties, which offer a novel natural approach to addressing inflammation-related diseases.