The Study On Extraction And Antitumor Activity Of Triterpenoids

Triterpenoids are the mainly nature bioactive compounds,the extraction yield of conventional extraction methods such as Organic solvent extraction is low.The extraction yield increased with the application of ultrasonic extraction(UE),microwave extraction(ME)and Ultrahigh pressure extraction(UPE)on the extraction of total triterpenoids,but UPE needs to be improved.The triterpenoids can significantly inhibit tunor,and its antitumor activity is always the study hot point,It is not enough to explore triterpenoids in our country,the drink comprised triterpenoids is few,it is promising to explore a Ganoderma Lcidum triterpenoid juice.Firstly,we combined pressure with temperature to extract total triterpenoids,applying high pressure extraction method,so it was more safety,the assay was conducted in High-pressure Reator Vessl,we compared the extraction yield of pressure extraction with UE and ME,in order to develop a new effective extraction method.Secondly,we selected Oleanolic acid and Ursolic acid as the material,we studied its antitumor activities on MGC803 and Caco-2 cell,then we studied its antioxidation activity on Ana-1 cells,which was harmed by H2O2.We studied the effect of structure difference on antitumor and antioxidation activities by testing the inhibition ratio of MGC803,Caco-2 cells and survival ratio of Ana-1 cells by MTT assays,in order to study whether the p53 protein played role in the MGC803 cell apoptosis,we tested the p53 expression by western blot.Lastly,we used the extracted triterpenoids to explore a juice,and studied its mouth feel,fiavour and stability,hoping to develop a drink that has a good fiavour and stability.The result showed that the high pressure extraction is an more effective extraction method,its the yield was 13.31mg/g in 30min,howerer,the highest yield of UE and ME were 10.08mg/g and 12.21mg/g,the yield of the high pressure extraction increased 30%and 10%,the result of response surface showed the optimical condition:87?,75%(v/v)ethanol,1Mpa,50ml/g,30min,and the pressure interacted with temperature.The result of HPLC showed Oleanolic acid and ursolic acid was stable in artificial simulated gastric/intestinal fluid.MTT assays showed Oleanolic acid and Ursolic acid could significantly inhibited the MGC803 and Caco-2 cells,enhanced the living ratio of Ana-1 cells,which harmed by H2O2,the IC50 concentration was 45?g/ml and30?g/ml for MGC803 cells,85?g/ml and 60?g/ml for Caco-2 cells.Ursolic acid had more strong effect.The western blot assays showed p53 protein in MGC803 cells was downregulated by Ursolic acid,the results showed that triterpendois,which have similar structure with Oleanolic acid and ursolic acid,have strong antitumor and antioxidation activities.The process research of triterpenoids juice showed sugar acid ratio affected the mouth feel,fruit juice affected flavour,and the hydrocolloid affected stability.After optimization,the content of drink is 25ml,0.1M citric acid,25ml,0.1M malic acid,150ml,1M sugar solvent,60ml apple fruit,30ml lemonade fruit,0.3g triterpenoids,the addition amount of eatable gun was 1%(the ratio of Xanthan gum and konjac gum is 2:1).