Optimization of extraction process and antioxidant mechanism of Polygonatum cyrtonema Hua polysaccharides
In this paper,a response surface methodology was employed to optimize the extraction process of Polygonatum cyrtonema Hua polysaccharides(PCP)by hot water based on single factor test,considering the parameters of ratio of raw material to water,extraction temperature and extrac-tion time.The PCP was isolated and purified to obtain water-washed polysaccharides.The results showed that the optimal extraction parameters were as follows:ratio of raw material to water of 1∶25,extraction temperature of 85 ℃,and extraction time of 2.0 h.On this basis,the antioxidant mechanism of PCP was investigated using oxidized low-density lipoprotein(Ox-LDL)-treated macro-phages.The MTT results indicated that PCP was not toxic to macrophages.Compared with the mod-el group,PCP significantly increased the levels of total antioxidant capacity(T-AOC)and the activities of glutathione peroxidase(GSH-Px)and superoxide dismutase(SOD),and decreased the content of malondialdehyde(MDA),indicating that PCP could enhance antioxidant effects in Ox-LDL-induced macrophages.Furthermore,Western Blot experiments showed that PCP could also significantly up-regulate the protein level of Nrf2 and down-regulate the protein expression of Keap1 in Ox-LDL-in-duced macrophages.These results revealed that PCP could regulate oxidative stress in macrophages induced by Ox-LDL,which might be related to the modulation of Keap1/Nrf2 signaling pathway.The study results may provide a theoretical basis for the further development and utilization of PCP.
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