Research Of The Constituent Characterization Of Phellinus Baumii Polyphenols And Its Antitumor Mechanism

Phellinus baumii,a precious traditional Chinese medicine,is named after a fungus that grows on mulberry trees.P.baumii is rich in active ingredients including polysaccharides,polyphenols,terpenoids,steroids,and etc.Modern pharmacological research has confirmed that it exerts many pharmacological effects like antioxidant,anti-inflammatory,immune regulation,antitumor,hypoglycemic and liver protection.Lots of studies have also shown that polyphenols show a wide range of pharmacological effects.It is of great significance to investigate the components of polyphenols in P.baumii and their antitumor activities for the development of antitumor drugs.In this study,the polyphenols of P.baumii(PBP)were extracted and purified,then the component identification,in vitro antitumor effect and the mechanism were investigated.The main results are as follows:(1)Polyphenols of P.baumii were extracted by ultrasonic-assisted ethanol reflux method.The crude extract was purified by a macroporous resin column and PBP with a high purity of 94.47± 4.10%was obtained.The UPLC-ESI-QTOF-MS technique and Peak View software were used to characterize the constituents of PBP.17 compounds like protocatechuic aldehyde,caffeic acid,osmundacetone were identified,their chemical structures were analyzed as well.Beyond that,the secondary mass spectra and the proposed fragmentation pathway of interfungin B,a representative component of PBP,were revealed.The results laid a significant foundation for the study of the pharmacological activities of P.baumii polyphenols.(2)The toxicity of PBP on human lung cancer cell A549,human colon cancer cell HCT116,human liver cancer cell HepG2,human cervical cancer cell HeLa,human bladder cancer cell T24 and human embryonic kidney cell HEK293 were determined by the CCK-8 method.The influence of PBP on the morphology of tumor cells was observed under a phase-contrast microscope.The results showed that PBP had a significant effect on inhibiting the proliferation of the above five tumor cells in vitro,in a concentration-and time-dependent manner.Among them,A549 and HCT116 cells were the most responsive to PBP with IC50 values of 49.1±0.5 μg/mL and 54.2±0.7μg/mL,respectively.On the contrary,PBP exhibited no significant toxic effect on normal cell HEK293 within the concentration of 101.3-101.9 μg/mL.PBP induced drastic and adverse morphological changes in A549 and HCT116 cells in a dose-dependent manner and they displayed typical features of cell apoptosis.(3)Analysis of cell apoptosis,cell cycle,reactive oxygen species level and mitochondrial membrane potential were performed using flow cytometry technique,on A549 and HCT116 cells treated by PBP.The results showed that PBP could induce cell apoptosis and S phase arrest of A549 and HCT116 cells.The level of reactive oxygen species of cells increased and the mitochondrial depolarization of A549 and HCT116 cells were induced significantly.These may be effective ways for PBP to inhibit the proliferation of A549 and HCT116 cells.These results provide a theoretical basis for the development of PBP as an antitumor drug.