Exploring the Potential Targets and Elucidating the Molecular Mechanisms Underlying the Anti-inflammatory Activity of <i>Plectranthus amboinicus</i> (Karpuravalli) in the Arachidonic Acid Metabolic Pathway by Network Pharmacology, Molecular Docking a

This work has been planned to predict the specific molecular targets, possible molecular mechanisms behind the anti-inflammatory potential of the phytoconstituents of Plectranthus amboinicus, specifically focusing on arachidonic acid metabolism using network pharmacological studies. Molecular modeling studies were done to determine the binding affinities of the ligands against the main anti-inflammatory targets. Also, in vitro anti-inflammatory study using RT-qPCR has been performed for validating the in silico results experimentally. From network pharmacological studies, overall eight nodes or targets named PTGS2, TNF, STAT3, RELA, PPARG, CYP2C19, CYP2C9 and ALOX5 were found to be the major anti-inflammatory targets of P. amboinicus. From GO and KEGG enrichment analysis, it can be concluded that the arachidonic acid metabolism pathway showed the highest fold enrichment and lowest [Formula: see text]-values. Molecular modeling studies showed that some of the phytoconstituents obtained from P. amboinicus showed moderate to good binding interactions with the respective proteins. Further two extracts were prepared using sequential extraction approach from the leaves of P. amboinicus, and the anti-inflammatory activity against the targeted gene obtained through in silico studies of the same was evaluated. For this purpose, at first, MTT assay was performed to determine the nontoxic concentrations of the prepared extracts in RAW 264.7 cells. Thereafter, RT-qPCR analysis has been done to determine the reduction of fold change in expression of COX-2 mRNA in the obtained cell supernatant. From in-vitro anti-inflammatory studies, it was evident that the prepared extracts showed promising inhibition against the COX-2 (PTGS2) enzyme. From this study, it can be concluded that P. amboinicus may exhibit anti-inflammatory activity by acting on the arachidonic acid metabolism pathway with specific interaction on COX-2 enzyme, and thus can be explored further for the development of selective COX-2 inhibitors.