| Objective:In recent years,anti-aging effect research of natural small molecule has been paid much attention at home and abroad and became a research hotspot.Licorice is rich in flavonoids,among the compounds reported to have biological activities are isolicoflavonol（Iso）,glabridin（Gla）,licochalcone A（LA）etc.They are widely used in anti-tumor,anti-oxidation,anti-inflammation,anti-diabetes,heart protection,nerve protection,enhance memory and other aspects.Therefore,through the experiment want to find out the chemical compound with anti-aging effect by analyzing the varities of compounds with biological activity in the flavonoids of licorice root.Methods:First of all,examine the bioactive of Iso,Gla,LA in hADSC by CCK-8 cell activity detection experiments.In hADSC,we investigated osteogenesis by Alizarin Red staining and adipogenesis by Oil Red O staining as well as ROS levels,SA-β-gal,relative telomere length and P16ink4a mRNA expression.RNA sequencing,western blotting and q RT-PCR examine the related signaling pathway.We also examine the glycolysis of hADSC.Treat with LA to aging C57BL/6 mice through morris water maze（MWM）test examine the spatial memory and cognition and examine cerebral blood flow（CBF）by Laser speckle contrast imaging,RNA-seq of Hipp on aging C57BL/6 mice to show that LA whether improve the spatial memory and cognition and by q RT-PCR detection the GAPDH,PFKP mRNA,use immunohistochemistry test SOX2 expression in Hipp and SVZ.Results:CCK-8 result showed that LA promote the proliferation of hADSC,osteogenesis and adipogenesis differentiation showed that LA improve the osteogenesis differentiation and inhibit adipogenesis differentiation,decrease the ROS levels,SA-β-gal,and P16ink4a mRNA relative expression,and also ameliorate the telomere shorter,showed that LA can inhibit the aging of hADSC.RNA-seq and western blot result showed that LA actiavate the PI3K/AKT signaling pathway in hADSC.The glycolysis level of hADSC was increase and mRNA expression of GAPDH and PFKP were promoted.MWM tests showed that LA improve the spatial memory and cognition,CBF result showed that LA have no effect on CBF and RNA-seq conducted on aging C57BL/6 mice showed that LA via the glycolysis pathway,as revealed by q RT-PCR detection of GAPDH,PFKP mRNA expression were increase in Hipp,and the number of SOX2 expression in Hipp was increased.Conclusion:Above all experiments results proved that LA can promote the proliferation of hADSC by activate PI3K/AKT signaling to promote the glycolytic pathway and inprove the spatial memory and cognition by improve the glycolytic pathway in aging C57BL/6 mice.This study showed that LA have the ability to anti-aging and provides a further important reference for improving the medicinal value of LA. |
