Polysaccharide is one of the main active ingredients in Lonicera japonica Thunb. (L. japonica). In this study, we examined the anti-aging activities of L. japonica polysaccharides (LJPs) and further explored the mechanisms. Polysaccharides from L. japonica including the crude LJP (CLJP) and the purified fraction (LJP-2-1) were characterized. The molecular weights of CLJP and LJP-2-1 were 1450&#
x202f;kDa and 1280&#
x202f;kDa, respectively. Meanwhile, CLJP was mainly composed of galacturonic acid (23.57&#
x202f;%), galactose (23.45&#
x202f;%) and arabinose (23.45&#
x202f;%). LJP-2-1 was mainly composed of galacturonic acid (51.25&#
x202f;%) and arabinose (22.89&#
x202f;%). In Caenorhabditis elegans (C. elegans), LJPs maximally prolonged mean lifespan by 13.97&#
x202f;%, promoted fitness with increased motility by 40.92&#
x202f;% and pharyngeal pumping by 25.72&#
x202f;%, and decreased lipofuscin accumulation by 38.9&#
x202f;% with intact body length and fecundity. Moreover, CLJP extended the mean lifespan of nematodes under oxidative and heat stress by 16.76&#
x202f;% and 14.05&#
x202f;% respectively by activating stress-related genes and the antioxidant system. Further, CLJP required DAF-16 to prolong the lifespan of nematodes. CLJP upregulated the expression of
daf-16 and its targeted downstream genes, including
sod-3,
gst-4 and
hsp-16.2. Moreover, nuclear accumulation of DAF-16 was promoted upon CLJP treatment. Together, our data uncover the role of LJPs in extending lifespan and healthspan through DAF-16.