Polysaccharide decolorization is a key determinant of polysaccharide structure. In this study, two purified Rehmannia glutinosa polysaccharides, RGP−1−A and RGP−2−A, were obtained after decolorization using the AB-8 macroporous resin and H₂O₂, respectively. RGP−1−A (molecular weight (Mw) = 18,964 Da) and RGP−2−A (Mw = 3305 Da) were acidic and neutral heteropolysaccharides, respectively, and were both polycrystalline in structure. FTIR analysis revealed that RGP−1−A was a sulfate polysaccharide, while RGP−2−A had no sulfate group. Experiments on IPEC-1 cells showed that RGPs alleviated oxidative stress by regulating the Nrf2/Keap1 pathway. These findings were confirmed by the upregulation of Nrf2, NQO1, and HO-1; the subsequent increase in the levels of antioxidant indicators (SOD, LDH, CAT, and MDA); and the restoration of mitochondrial membrane potential. Overall, the antioxidant capacity of RGP−1−A was significantly higher than that of RGP−2−A. These results suggest that RGPs may be a potential natural antioxidant and could be developed into functional foods.

Laying hens over 75 weeks of age commonly show great declines in immunity and production performance. It is unclear whether these declines can be relieved by supplementing with ascorbic acid (AA) in feed. Two trials were conducted to investigate the synthesis and metabolism of AA in layers of different ages and the effects of dietary supplemental AA on the performance and the immune and antioxidant statuses of 78 weeks old hens. In Exp. 1, equal numbers (24 hens) of 35 weeks old (Young) and 75 weeks old (Old) layers were fed the same diet without AA supplementation for 4 weeks. In Exp. 2, 360 healthy 78 weeks old laying hens were randomly assigned to 4 treatments (basal diet supplemented with 0, 0.25, 0.5, or 1 g AA/kg diet) in an 8-week feeding trial. The old hens tended to have decreased L-gulonolactone oxidase (GLO) synthase activity in the kidney and liver than that of the young hens (P = 0.07 and P = 0.05, respectively). Compared with the young hens, the old hens had lower hepatic antioxidant capacity allowing for the lower thioredoxin (TXN), thioredoxin reductase (TXNR) and cytochrome b5 reductase (CYB5R) gene expression (P < 0.05), whereas increased sodium-dependent vitamin C transporter (SVCT) 1 expression levels in the ileum and kidney and enhanced splenic and hepatic AA concentrations (P < 0.05). Dietary supplementation with AA significantly decreased GLO enzyme activity but increased splenic AA concentration and anti-bovine serum albumin IgG levels (P < 0.05) and tended to increase CD4+ T lymphocyte numbers (P = 0.06) in serum. Supplementation of 0.25 g AA/kg diet significantly increased hepatic total antioxidant capacity (T-AOC, P < 0.05) relative to the control group. Laying hens could synthesize AA in both the kidney and the liver, though the GLO enzyme activities were 100 times greater in kidneys than in livers. The old laying hens had greater absorption and reabsorption capacity and higher AA retention in some tissues that did the young hens. Dietary supplementation of AA can improve the health of old layers by enhancing immunity and antioxidant capacity.