Lactuca indica (Compositae) is an edible wild vegetable, used as a folk medicine in anti-inflammatory, antibacterial, and other medications in Asia. This is the first scientific evaluation of the chemopreventive therapeutic properties of L. indica using five antioxidation assay systems. An extract from L. indica was found to possess significant free radical scavenging activity, effectively protecting phix174 supercoiled DNA against strand cleavage and reducing oxidative stress in human promyelocytic leukemia HL-60 cells. Moreover, extracts of L. indica almost totally inhibited nitric oxide production and the mRNA expression of inducible nitric oxide synthase, at a dosage of 100 microg/mL, in LPS-stimulated macrophage RAW264.7 cells. Bioactivity-guided chromatographic fractionation and metabolite profiling coupled with spectroscopic analyses revealed that the six phenolic compounds, that is, protocatechulic acid (1), methyl p-hydroxybenzoate (2), caffeic acid (3), 3,5-dicaffeoylquinic acid (4), luteolin 7-O-beta-glucopyranoside (5), and quercetin 3-O-beta-glucopyranoside (6), are the major antioxidative constituents in the L. indica extract.