Bioseparation And Anti-Aging Activity Of Hydroxytyrosol-1-Glucopyranoside

Hydroxytyrosol-1-glucopyranoside(HTG)is a kind of natural phenylethanol glycosides in the plant Sargentodoxa cuneata(Oliv.).Its aglycon hydroxytyrosol has anti-aging effect,but the anti-aging effect and mechanism of HTG have not been reported.HTG is usually obtained by plant extraction,which has some problems such as complex composition,low extraction efficiency and long cycle.However,bioconversion production of HTG is a green and sustainable alternative.Therefore,in this study,hydroxytyrosol was biotransformed into HTG,which was separated and purified by macroporous resin and preparative HPLC.Subsequently,D-galactose(D-gal)induced aging model was established to investigate its anti-aging activity and possible mechanism in vivo and in vitro.Bioconversion to HTG using hydroxytyrosol as a substrate.When 1 g/L hydroxytyrosol was added into the flask,the titer for 48 h was 1.81 g /L.In the fed-batch fermentation,4 g/L hydroxytyrosol was added at 12-36 h with a titer of 6.80 g/L at 48 h.Subsequently,SP825 L macroporous resin was screened from 5 different macroporous resins,which could efficiently separate HTG.After purification by SP825 L resin,the purity of the samples was 81.71% ± 1.63% and the recovery was 82.43% ± 2.31%.Further purification using preparative liquid chromatography results in HTG with a purity of more than 95%.To explore the protective effect of HTG on D-gal induced HFF cell senescence and its mechanism.HTG can improve the survival rate of HFF cells after D-gal induced aging,reduce the increase of SA-β-Gal activity in cells,reduce the m RNA expression levels of P53 and P21,reduce the apoptosis rate and the expression of apoptotic protein,and alleviate DNA damage.It can reduce the production of intracellular ROS,improve antioxidant capacity and reduce the occurrence of oxidative stress.It can reduce the m RNA expression of SASP factor and reduce the occurrence of intracellular inflammation.Its possible mechanism of action is to improve the metabolic imbalance caused by D-gal,enhance the antioxidant defense ability of cells,maintain the redox dynamic balance,and reduce the cell oxidative damage induced by D-gal by activating AMPK and Nrf2 signaling pathways.To explore the protective effect of HTG on D-gal induced aging in mice and its mechanism.HTG can increase the body weight and organ index of D-gal induced aging mice,improve the skin and organ tissue damage,restore abnormal blood lipid index,and has a certain protective effect on aging mice.It can reduce MDA content in liver and kidney,increase SOD and CAT activity,and improve antioxidant capacity of mice.It can reduce the level of inflammatory cytokines,inhibit the expression of NF-κB signaling pathway protein,and reduce the occurrence of inflammation in vivo.Its possible mechanism is to improve metabolic imbalance,enhance antioxidant defense ability,maintain redox dynamic balance,and reduce oxidative damage induced by D-gal by activating AMPK and Nrf2 signaling pathways.This study established a bioseparation method for HTG,proved the anti-aging effect of HTG in vivo and in vitro,studied the possible mechanism of action,and laid a foundation for its application in health care products,cosmetics and other fields.