The aim of this study is to investigate hypolipidemic and antioxidant effects of Pine needle polysaccharide (PNP) from Pinus massoniana in high-fat diet (HFD)-induced mice. PNP could significantly improve the serum lipid levels (total cholesterol, triacylglycerols, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol), enhance the antioxidant enzymes levels (total antioxidant capability, superoxide dismutase, glutathione peroxidase, catalase), and decrease malondialdehyde (MDA) content in HFD-induced mice. PNP exhibited distinct antioxidant ability on the superoxide anions, 1, 1-diphenyl-2-picrylhydrazyl (DPPH) and ferric ion reducing antioxidant power (FRAP) in vitro. The average molecular weight (Mw) of PNP was 6.17 × 10⁵ Da, and mainly of fucose, arabinose, galactose, glucose, galacturonic acid. These results suggested that PNP might be used as functional foods and natural drugs in enhancing antioxidant ability and alleviating the hyperlipidemia.