Background: Hypericin is an anthraquinone derivative of Hypericum hookerianum, a shrub from Palni hills of Southern India which possess various medicinal values. Aim: The present study was aimed to evaluate the anti-inflammatory activity of hypericin using RAW 264.7 macrophage cell line. Materials and Methods: The anti inflammatory property was determined by assessing the inhibitory action on lipopolysaccharide (LPS) stimulated nitric oxide (NO) production and pro inflammatory mediators/cytokines. RAW 264.7 macrophages cell line was used as in vitro inflammatory model. Cytotoxicity was evaluated using methylthiazolyldiphenyl tetrazolium bromide assay and NO estimation was carried out using Griess method. Gene expression of pro—inflammatory cytokines such as inducible nitric oxide synthase, cyclooxygenase 2, tumor necrosis factor α, interleukin—1 β (IL—1 β), and IL—6 was also evaluated. Results: The results exhibited that hypericin significantly suppressed LPS induced NO production with a concomitant decrease in the levels of pro—inflammatory cytokines. Conclusion: The study demonstrates the potential of hypericin as an effective anti—inflammatory agent.