Saccharin and sucralose are worldwide used tabletop non-caloric artificial sweeteners (NAS), and although they are considered metabolically inert and safe for consumption, recent studies observed an NAS-mediated modulation of the intestinal microbiota, which may result in the development of intestinal inflammatory diseases. The present study investigated the effects of NAS alone and in combination with dextran sodium sulfate (DSS), a chemical known to induce gut inflammation, on parameters of overall fitness including longevity and climbing ability of female Drosophila melanogaster. DSS-exposed flies were subjected to saccharin or sucralose, and their effects on survival and the gene expression levels of different antimicrobial peptides (AMPs) were assessed. Saccharin supplementation significantly increased the lifespan of D. melanogaster, while DSS significantly decreased the survival in a dose-dependent manner. Additionally, DSS exposure of flies resulted in a trend toward an up-regulation of the mRNA levels of the AMPs, drosocin and metchnikowin. This effect was more pronounced in the NAS-supplemented group reaching significance. This may indicate that NAS supplementation improves intestinal immune reactions during an inflammatory metabolic state.