Studies On Purification And Activities Of Anti-prostate Cancer PC-3 Cell In Vitro For The Pomegranate Peel Punicalagin In Xinjiang

Objective:1.To establish the purification process and amplification test laboratory for Punicalagin of Pomegranate Peel.2.To study the inhibitory effect of Pomegranate Peel Punicalagin and Pomegranate Peel Polyphenols(the content of Punicalagin was 40.70%)on the proliferation of human prostate cancer PC-3 cell and its possible mechanism.3.To determine the median lethal dose of Pomegranate Peel Punicalagin.Methods:1.The purification process of Pomegranate Peel Punicalagin was screened by single factor analysis with the content of Punicalagin as index.2.Using cell morphological observation and MTT assay determination of different concentrations of Pomegranate Peel Punicalagin and Pomegranate Peel Polyphenols impact on PC-3 cells proliferation,different concentrations of Pomegranate Peel Punicalagin and Pomegranate Peel Polyphenols after treated with 48 h PC-3 cell cycle phase change and apoptosis by flow cytometry,the results were compared,and with the positive control drug flutamide comparison.3.To use the single dose toxicity test to determine median lethal dose of Pomegranate Peel Punicalagin.Results:1.The purification process of Pomegranate Peel Punicalagin was as follows:Pomegranate peel crushing after 80 mesh sieve,according to the material liquid 1:24,add 60%ethanol,leaching for 30 min at room temperature,gauze filtration,the filtrate decompression to concentrate without alcohol flavor,Diluted with distilled water to 20.8 mg/mL,1.2 mol/L HCL mix the pH to 2,choose HPD500 resin,with 2 BV/h dynamic adsorption speed sample 5 BV,let stand for 12 h,7 BV with distilled water and 5 BV 5%ethanol in removing impurity,6 BV 10%ethanol elution at 4 BV/h speed,collect 10%ethanol elution fluid,the reduced pressure concentration and freeze drying.2.Pomegranate Peel Punicalagin and Pomegranate Peel Polyphenols significantly inhibited the cell proliferation of PC-3 cells in dose and time dependent manners.The effect of Pomegranate Peel Punicalagin was better than the effect of Pomegranate Peel Polyphenols(P<0.05).There was no significant difference between Pomegranate Peel Punicalagin and Pomegranate Peel Polyphenols(P>0.05).Pomegranate Peel Punicalagin and Pomegranate Peel Polyphenols obviously induce the apoptosis of PC-3 cells in dose dependent manners.When the mass concentration were 400 and 800 μg/mL,Pomegranate Peel Punicalagin group of early apoptosis rate is superior to the Pomegranate Peel Polyphenols,there was significant difference between the effect of inducing apoptosis of Pomegranate Peel Punicalagin and Pomegranate Peel Polyphenols(P<0.05).When the mass concentration were 200 and 800 μg/mL,there was no significant difference between the effect of inducing apoptosis of Pomegranate Peel Punicalagin and flutamide(P>0.05).The Pomegranate Peel Punicalagin、Pomegranate Peel Polyphenols and flutamide make G0/G1 phase cells showed a trend of increasing the proportion,S stage cells ratio decreased significantly,when the mass concentration of 400、800 μg/mL,compared with negative control group,the difference was statistically significant(P<0.05).3.Pomegranate Peel Punicalagin median lethal dose(LD50)is 2615.6 mg/kg,the 95%confidence limits the range of 2336.5-2336.5 mg/kg.Conclusions:1.After macroporous adsorption resin purification of Punicalagin in Pomegranate Peel Punicalagin content increased significantly,this method is simple,in order to further enlarge production laid the foundation experiment.2.Pomegranate Peel Punicalagin in vitro significantly inhibit prostate cancer PC-3 cell proliferation,inducing apoptosis and cell cycle,to block the PC-3 cells in G0/G1 phase,the cell proliferation cycle extended,proliferation at a slower pace.3.With the increase of extract with pomegranate contents,the median lethal dose is reduced,its toxicity may be associated with the content of Pomegranate Peel Punicalagin.