Inhibitory Effects And Mechanisms Of Extracts From Distiller’s Grains On HepG2 Cells

Objective: To preliminarily explore components of extracts from distillers’ grains and its inhibitory effects and mechanisms on HepG2 cells.Methods: The components of extracts from distillers’ grains were analyzed by LC-MS.RTCA was used to detect the inhibitory effects of different concentrations of extracts from Distiller’s grains on HepG2 cells.After stimulating for 24 h,the cell cycle distribution of HepG2 cells was detected by flow cytometry;Real-Time qPCR and Western Blot was used to detect the expression of cyclic related molecule and their corresponding dependent kinases;Transwell assay was used to detect the migration and invasion ability;Flow cytometry was used to detect the changes of apoptosis rate of HepG2 cells;Real-Time qPCR and Western Blot were used to detect the expression of apoptosis-related marker molecules;The expression of PI3K-AKT pathway related molecules was detected by Western Blot.Results: 43 substances in extracts from distillers’ grains were determined by LC-MS.Extracts from Distiller’s grains with a concentration of 24.9μg/ml had little effect on the proliferation of HepG2 cells(p>0.05).Extracts from Distiller’s grains with a concentration of 49.5μg/ml had little effect on the proliferation of HepG2 cells at 12h(p>0.05).In addition,with the increase of concentration and stimulation time,compared with the control group 0ug/ml,the proliferation ability of HepG2 cells in the treatment group was significantly inhibited(p<0.05),and its IC50 of 24 h treatment is 75.5 μg/ml.The invasion and migration ability of HepG2 cells in treatment group was significantly inhibited,and the number of inhibited HepG2 cells increased with the concentration of distiller’s grains(p<0.05).with the increase of concentration ofextracts from Distiller’s grains,the proportion of S phase of cell cyle in HepG2 cells increased gradually,and the expression of CyclinA and its kinase CDK2 decreased gradually.The expression levels of proapoptotic molecules Bax,CYTC,Cleaved-Caspase-9 and Cleaved-Caspase-3 in the treatment group were gradually increased compared with the control group(P< 0.05),while the expression of anti-apoptotic molecules Bcl-2,Caspase-9 and Caspase-3 decreased gradually(p<0.05);The expression of p-PI3 K and p-AKT in PI3K-AKT pathway decreased gradually(p<0.05),and there was no significant change in PI3 K and t-AKT levels(p>0.05).Conclusion:Extracts from distillers’ grains contains multiple active substances.Extracts from Distiller’s grains can inhibit the proliferation of HepG2 cells in time-dependent and dose-dependent manners.Extracts from Distiller’s grains can inhibit the migration of HepG2 cells in a concentration-dependent manner;Extracts from Distiller’s grains arrestHepG2 cell in the S phase of the cell cycle by inhabiting the expression of CyclinA and CDK2.Extracts from Distiller’s grains promote apoptosis of HepG2 cells by inhibiting the PI3K-AKT pathway and activating the mitochondrial pathway that mediates apoptosis.