| Modern research shows that Agrocybe aegerita polysaccharide(AAP)has antioxidant and anti-aging and other physiological functions,which is one of the important sources of active polysaccharides.Resveratrol(RES)is a natural antioxidant in plants.A large number of in vitro and in vivo studies have shown that RES also plays an important role in anti-aging effects.The synergistic anti-aging effect on AAP with RES has a good application prospect.In this study,the optimum extraction process was determined by single factor and orthogonal test.The antioxidant effects of AAP and RES in vitro and in vivo were studied,and the synergistic anti-aging effect was tested.1.The extraction temperature,extraction time and ratio of material to liquid were selected as the influencing factors by using hot water extraction AAP method.Orthogonal test was designed.The optimum extraction was determined by using glucose content as the determination index.The best extraction conditions were as follows:extraction time 3.0h,extraction temperature 95℃,ratio of material to liquid 1:40,the polysaccharide yield was 3.18%.2.The antioxidant capacity of AAP and RES as well as combination of them were studied by DPPH,ABTS,hydroxyl radical and reducing power.AAP has a good scavenging effect on free radicals,but also has a strong reducing power.Its antioxidant activity increased with the increase of concentration.The IC50 of AAP and RES compound was lower than that of single component,and the CI was less than 1,indicating that the compound showed significant synergistic effect in the evaluation system.3.The model of MRC-5 cell senescence was induced by H2O2.MTT assay was used to detect the effects of AAP and RES on the MRC-5 cells viability.The telomerase activity was detected by TRAP;The GSH content was detected by colorimetry;Cell cycle was analyzed by flow cytometry.The results showed that,1)After the intervention of AAP and RES,the aging cell metabolic activity increased with the concentration increasing,and the cell survival rate of the combination group increased significantly;2)GSH and telomerase activity in senescent cells were significantly lower than those in the blank group.Compared with the model group,they were significantly increased in all the drug intervention groups;The GSH and telomerase activity of the compound group were better than that of the single component,which showed that the synergistic effect of anti aging.3)The results of cell cycle showed that AAP and RES could delay the cell arrest in G1 phase and promote cell proliferation.The combination could significantly increase the proportion of S phase cells and promote the cell proliferation.4.D-gal was injected subcutaneously to establish the model of aging mice.The rats were treated with AAP and RES for 8 weeks.The changes of body weight were measured.The effects of aging model on blood index and biochemical indexes were studied.The colorimetric method was used to determine the activity of SOD,GSH-Px,CAT,T-AOC and MDA.The histopathological changes of liver were observed by HE staining.The results showed that,1)The ALT and AST level of aging mice serum decreased significantly,but there were no significant differences in body weight,blood cells and blood lipids index;2)Compared with the blank group,the activity of SOD,T-AOC,GSH-Px and CAT in the liver tissue of the male and female mice were significantly decreased,and the content of MDA was increased.Compared with the aging model group,the AAP could significantly increase the antioxidant activity Enzyme activity,while reducing MDA content.The combination showed synergistic anti-aging effects,and the male mice had more anti-aging effects.3)The normal tissue structure of liver tissue of HE mice was decreased,and the damage degree of other drug intervention group was lighter than that of model group,and the combination had protective effect on the liver tissue of aging mice. |
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