Optimization Of Culture Conditions Of Cordyceps Kyushuensis And Its Antioxidative And Anticancer Activity In Vitro

Objective: In order to study the relationship between the biomass and the contents ofthe relative bioactive substances in submerged culture, the antioxidation and anticanceractivities in vivo. The optimization of the biomass in liquid culture conditions ofCordyceps kyushuensis were researched. The antioxidation, anticancer activities andpharmacological mechanism in vivo were also researched to offer reference to find newantitumor compounds.Methods: Optimal cultivation conditions of biomass were researched by Single Factorand Box-Behnken test, the dry weight of mycelia were researched by dry weighingmethod. The phenol-sulphate acid method and3,5-dinitrosalicylic acid method were usedto quantify the intracellular polysaccharide and extracellular polysaccharide contents. Thepotassium periodate oxidation method was used to quantify the extracellular mannitol andintracellular mannitol contents of those samples. The antioxidation of the total antioxidantcapacity (T-AOC), atalase activity (CAT) and glutathione peroxidase (GSH-PX) wasdetermined by antioxidant trizol. The anticancer extract samples were prepared andfollowed identifying the components. In vitro, cell growth inhibitory effects of theseextracts against human hepato carcinoma cell HepG2, human gastric carcinoma cellBGC823, human cervical cell HeLa, human gastric tumor cells AGS and theircytotoxicity to the human embryonic lung fibroblast cell HELF and human gastricepithelium normal cells GES1were evaluated by MTT assay in time and dose. Themorphological change of cells was observed under light microscope and fluorescencemicroscope with AO stained after treatment with samples. The flow ctometry was used todetect cell cycle distribution.Results: The best liquid medium for the biomass proved by the Box-Behnkenexperiment added with Zn in the medium was glucose6.04%, peptone2.77%, andKH2PO40.1%, MgSO40.29%g/L, ZnSO4200mg/L, liquid medium volume100ml/250mL, pH value6.0. The biomass of mycelia from the medium added with Zn was 2.97times weigher than that from basal medium without Zn. The total antioxidantcapacity (T-AOC), atalase activity (CAT) and glutathione peroxidase (GSH-PX) ofintracellular extracts of Cordyceps kyushuensis cultured in Zinc-Rich medium wereincreased significantly. After analysis, it were verified that ergosterol and others may beincluded in the ethyl acetate and chloroform extracts. The inhibiting capacity of differentscells were not the same in diffenent time and dose by MTT. There were little cytotoxicityto the normal cells by the samples. In anti-cancer mechanism research, apoptosismorphological changes about chromatic agglutination and nuclear condensation weredetected by light microscope and fluorescence microscope with AO staining aftertreatment with ethyl acetate extract. The number of the tiny fragments increased with theconcentration of ethyl acetate extract, and cell cycle for HepG2and AGS were blocked inG1or S period.Conclusion: It is important to optimize the culture conditions for the higher biomassand bioactive compounds. And Cordyceps kyushuensis was verified that it has theantioxidation and anticancer activities.