| In the planting process of P.lactiflora,its above-ground parts are often abandoned,causing environmental pollution and waste of resources.The P.lactiflora flower is one of the above-ground parts of P.lactiflora.It is non-toxic,edible,and has high ornamental and therapeutic value.Microbial fermentation can enhance the application value of Chinese herbal medicines.In this study,modern microbial fermentation technology was used to perform liquid fermentation of P.lactiflora flowers with three different yeasts(Saccharomycopsis fibuligera,Saccharomyces cerevisiae,and BY4741 Saccharomyces cerevisiae).Furthermore,the effects of different yeast fermentation on the active ingredients and physiological activities of P.lactiflora flowers were evaluated through the analysis of antioxidant activity,anti-aging activity,and active ingredients,which provided a theoretical basis for the development of P.lactiflora flowers products,and also provided a new way for the development and utilization of Chinese herbal medicine.1.The effect of yeast fermentation on the antioxidant activity of P.lactiflora flowersExplore the effect of yeast fermentation on the antioxidant activity of P.lactiflora flower through a variety of in vitro antioxidant activities.The results showed that the content of total phenols and total flavonoids,DPPH free radical elimination activity,total antioxidant activity,total reduction capacity and protein oxidative damage protection activity of the P.lactiflora flower S.fibuligera fermentation liquid(SF)were significantly higher than those of the unfermented extract(NF),BY4741 S.cerevisiae fermentation liquid(BS),and S.cerevisiae fermentation liquid(SC).The contents of total phenols and total flavonoids of SF were 186.33± 0.44 mg/g and 39.04 ± 0.45 mg/g,respectively;When the concentration is lmg/mL,the protective effect of protein oxidative damage is 69.1%;when the concentration is 0.1 mg/mL,the total antioxidant activity and total reducing capacity of SF are increased by 69.6%and 101.65%,respectively,compared with NF.It shows that yeast fermentation can improve the antioxidant of P.lactiflora flowers,and the fermentation of S.fibuligera can increase the antioxidant activity of P.lactiflora flowers the most2.Effect of yeasts fermentation on the anti-aging activity of P.lactiflora flowers H2O2 induced NIH3T3 cell senescence model was used to evaluate the anti-aging activity of the S.fibuligera fermentation liquid of P.lactiflora flowers.The result shows that the three kinds of yeast fermentation liquid and unfermented extracts of P.lactiflora flowers can improve the reduction of cell viability induced by H2O2.The cell viability of the SF treatment group is the strongest.When the concentration is 800 μg/mL,the cell survival rate reaches 95.67%;in the cell morphology analysis,the cell morphology of the SF treatment group was stretched compared to the H2O2 treatment group,and the degree of stretch was significantly higher than that of the NF treatment group;in the analysis of ROS(reactive oxygen species)content and cell death,the ROS content and the number of cell deaths in the SF treatment group were significantly lower than the H2O2 treatment group and the NF treatment group;in the MMP(Mitochondrial membrane potential)analysis,the MMP intensity of the SF treatment group was significantly higher than that of the NF treatment group;in the apoptosis and cell cycle analysis,the number of apoptotic cells and the number of cells in the G1/GO phase of the SF treatment group were significantly lower than that of the NF and H2O2 treatment group.This shows that the anti-aging activity of SF is significantly higher than that of NF,and SF and NF reduce the damage of ROS to the inner mitochondrial membrane by reducing the content of ROS,thereby protecting the nucleus,and preventing cell apoptosis and cell cycle arrest.3.Antioxidant and anticholinesterase activity of different extracts from the S.fibuligera fermentation liquid of P.lactiflora flowersAnalyze the antioxidant activity and anticholinesterase activity of SF and its extracts.The results showed that the antioxidant activity of the ethyl acetate layer and n-butanol layer of the S.fibuligera fermentation liquid of P.lactiflora was significantly higher than that of the S fibul igera fermentation liquid of P.lactiflora flowers;among the anti-acetyl/butyrylcholinesterase activities,the cholinesterase inhibitory activity of the ethyl acetate layer and n-butanol layer of the S.fibuligera fermentation liquid of P.lactiflora flowers was significantly higher than that of the S.fibuligera fermentation liquid of P.lactiflora flowers,indicating that the main active part of acetyl/butyrylcholinesterase are ethyl acetate layer and n-butanol layer.4.Changes of active compounds before and after S.fibuligera fermentation of P.lactiflora flowersChanges of active compounds before and after S.fibuligera fermentation of P.lactiflora flowers were analyzed by HPLC-MS.The results showed that 8 polyphenols and 2 monoterpene glycosides were detected and identified in SF,and 10 polyphenols and 3 monoterpene glycosides were detected and identified in NF.And the content of gallic acid,astragaloside and quercetin-3-O-glucoside is significantly higher than that of NF;the 1,2,3,4,6-O-pentagalloyl glucose,quercetin-3-O-β-D-glucose-7-O-β-D-gentioside and oxypaeoniflorin detected in NF,but these compounds were not detected in SF;and in the HPLC-MS analysis of DPPH pre-column pretreatment,it was determined that gallic acid and quercetin were the main active compounds to scavenging DPPH free radicals.5.Molecular docking analysis of anti-oxidation and anti-aging of all active compounds of P.lactiflora flowersThe molecular docking of active compounds in P.lactiflora flowers and anti-oxidant or anti-aging target enzymes in P.lactiflora flowers were studied The results show that the active compounds of P.lactiflora flowers can interact well with the target protein.Among them,benzoyloxypaeoniflorin has the strongest interaction with the four enzymes,and the molecular docking scores with catalase,glutathione synthase,tyrosinase and SIRT 1 are-14.0989 Kcal/mol,-11.2059 Kcal/mol,-11.1193 Kcal/mol and-12.4587 Kcal/mol.It shows that benzoyloxypaeoniflorin may be the main active compounds of anti-oxidation and anti-aging in vivo.In summary,fermentation of S.fibuligera can significantly increase the content of total phenols,total flavonoids and anti-oxidation,anti-aging,and anti-cholinesterase activities of P.lactiflora flowers,the main compounds increase after fermentation are gallic acid,astragalin and quercetin-3-O-β-D-glucoside.Benzoyloxypaeoniflorin may be the main active substance of P.lactiflora flowers for anti-oxidation and anti-aging in vivo. |
