Antioxdant And Antiproliferative Activity Of Polyphenols Extract From Morchella Angusticepes Peck

In this paper,bioactive substances of Morchella angusticepes Peck was extracted by in vitro digestion.The polyphenols content and antioxidant activity by oxygen radical absorption capacity(ORAC)were determined.Antiproliferation and cytotoxicity were measured in hepatoma cell line HepG2 and colon cancer cell line Caco-2.Antioxidant activity was evaluated by cellular antioxidant activity(CAA)assay in HepG2 cells,it can reflect the antioxidant activity in cellular.In addition,in vitro digestion of Morchella angusticepes Peck showed antiproliferation and cellular antioxidant activity,which provides theoretical basis for the further development and utilization of morels.The HepG2 and Caco-2 were incubated with the polyphenols extracted from Morchella angusticepes Peck by the extraction of acetone solvent.And the content of enzyme activity and protein in HepG2 cells were determined to prove the mechanism of antioxidation and antiproliferative effect of polyphenols extracted from Morchella angusticepes Peck.The main conclusions are as follows.(1)After the simulation of gastrointestinal digestion,polyphenol and the value of ORAC were increased significantly.Result of the antiproliferative activity showed that HepG2 cells and Caco-2 cells were significant inhibited.The in vitro simulated gastric digestion could not inhibit the proliferation of HepG2 cells,but the in vitro simulated intestinal digestion had a strong inhibitory effect on HepG2 cells.In the case of Caco-2 cells,the proliferation of Caco-2 cells was stronger than that of gastric acid group and stomach blank group,stomach 0 h,gastric group,intestinal group and intestinal blank group were all have an significant effect on Caco-2 cells.CAA results showed that the samples showed prooxidation in gastric digestion and the samples showed antioxidant in intestinal digestion,the CAA value of intestinal digestion 0.5 h was(52.32 ± 7.61 ?mol QE/g DW).The CAA value of intestinal digestion 0.5 h was bigger than that of intestinal blank group 0.5 h(32.06 ± 3.05 ?mol QE/g DW).(2)The free and bound polyphenols extract from Morchella angusticepes Peck show antioxidant activity in HepG2 cells,the CAA value of them were 26.60±5.35 ?mol QE/g DW?15.03±2.15 ?mol QE/g DW,respectively,and with the concentration increases,the more obvious antioxidant activity of free and bound polyphenols extract.Antioxidant activity may be related to the antioxidative signaling pathway of Nrf2-Keap1-ARE,which is characterized by promoting the expression of Nrf2 protein,Nrf2 entering the nucleus and initiating the transcription of the lower phase II detoxification enzyme and cytogenetic protein gene of ARE,then,to protect the normal function of cells(3)The free polyphenols extract from Morchella angusticepes Peck can effectively inhibit the proliferation of HepG2 and Caco-2 and there is a significant dose-effect relationship,with the increase of the concentration,the inhibitory were more obvious.The inhibitory effect on HepG2 was better than that on Caco-2,and the antiproliferative effect of the bound polyphenols extract from Morchella angusticepes Peck is not significantly.(4)The free polyphenols extract from Morchella angusticepes Peck can inhibit cells proliferation and promote apoptosis,which maybe regard to activate the p38/MAPK signal pathway in HepG2 cells.The expression of p21,p-ASK1 and p-p38 in HepG2 cells were promoted and the expression of p-p53,CDK4,Cyclin D1,PCNA and TRAF-2 protein were inhibited by the free polyphenols extract from Morchella angusticepes Peck,effect the normal cell cycle and inhibit cell proliferation.The expression of Bax and Caspase-3 protein in HepG2 cells were promoted and the expression of Bcl-2 protein was inhibited by the free polyphenols extract from Morchella angusticepes Peck,and then induced apoptosis.