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Glutaredoxin (Grx) is a polypeptide with low molecular weight, which has been extracted from buckwheat and has been suggested to have multiple functions revolving around oxidative stress responses and cell signaling. Here, we report the antioxidant activity of recombinant buckwheat Grx (rbGrx) to reduce aging effects in Caenorhabditis elegans (C. elegans) as well as the mechanism involved. Our results showed that rbGrx beneficially affected the health span of C. elegans, including pharyngeal-pumping rate, locomotion, and lipofuscin accumulation. Furthermore, stress assay showed that rbGrx could extend the lifespan under both oxidative and heat stress. Further studies indicated that the longevity-extending effects of rbGrx could be attributed to its in vitro and in vivo antioxidant activities. After treatment with rbGrx, SOD activity, CAT activity, GSH content, and GSH/GSSG ratio were increased, while MDA content was decreased, which led to low intracellular levels of reactive oxygen species in C. elegans. Moreover, rbGrx up-regulated hsf-1 mRNA level and could not expand the lifespan of the hsf-1 mutant C. elegans (sy441); however, this had no effect on the transcription of daf-16 and skn-1 and could expand the lifespan of both daf-16 and skn-1 mutants. These results suggested dependency of the rbGrx effect on the heat shock transcription factor (HSF-1) and independency on both DAF-16 and SKN-1. In summary, our results demonstrated the anti-aging activity of rbGrx, which increased resistance to cellular stress and improved the health span of C. elegans. These results are very important for the use of rbGrx in anti-aging research.rbGrx improves healthspan and resistance to heat and oxidation stress.Benefit effects of rbGrx are due to upregulation hsf-1.This is the first time to evaluate anti-aging using recombinant glutaredoxin derived from buckwheat.rbGrx may promote healthy life as a dietary supplement.