Comparative Analysis of the Impact of Different Ecotypes on In Vitro Anti-Inflammatory Activity of Ethanolic Extracts of Moringa oleifera Leaves

Moringa oleifera (MO) is traditionally used to mitigate inflammatory-mediated disorders; however, the influence of ecotypic variation on its anti-inflammatory activity remains poorly understood. In this study, we compared the phytochemical composition and anti-inflammatory activity of ethanolic extracts obtained from fresh and dried leaves of four MO ecotypes (India, Paraguay, Mozambique, and Pakistan), all grown under the same outdoor conditions, as well as two commercial powders (Just Moringa and WISSA), using LPS-stimulated RAW 264.7 macrophages. Extracts from fresh leaves were 19–43% more cytotoxic than those from dried leaves, depending on the ecotype, likely due to higher cyanogenic glycoside content. Extracts from the India and Paraguay ecotypes, characterized by high levels of quercetin derivatives and caffeic acids, as well as Just Moringa, enriched in kaempferol derivatives, significantly inhibited LPS-induced nitric oxide (NO) production (p < 0.05). Just Moringa and Paraguay extracts also reduced iNOS gene expression (p < 0.05 and p < 0.01, respectively), whereas only the Paraguay extract decreased iNOS protein levels (p < 0.05). In contrast, quercetin-3-O-glucoside and rutin showed significant effects only at concentrations approximately 100-fold higher than those present in the extracts, indicating that the phytocomplex displays greater bioactivity than individual compounds. Overall, these results demonstrate that ecotypic variation strongly affects the polyphenolic composition and anti-inflammatory properties of MO leaves, highlighting the importance of reporting both origin and phytochemical composition in MO-based products.