| Prostate cancer is one of the most common malignant tumors among men.In the world,the incidence of prostate cancer ranks second among all malignant tumors in men,which seriously endangers the health of male patients.At present,in the process of cancer prevention and treatment in China,traditional Chinese medicine plays an important role in improving immunity,inhibiting the proliferation and metastasis of cancer cells,promoting apoptosis and reversing multidrug resistance.Ziziphus jujuba Mill.,is the fruit of jujube tree of the genus Rhamnoides.Traditional Chinese medicine considers that jujube is a kind of homologous medicinal material with edible and medicine food function,which has extremely rich nutritional and health care value.There are many varieties of Ziziphus jujuba Mill.in China,and Ziziphus jujuba Mill.resources are abundant.As a uinique variety in China,Jinsi jujube(Jinsixiaozao)has excellent flavor and:invaluable medicine effect.Laoling is known as"the hometown of Jinsi jujube in China".Ziziphus jujuba Mill.has many pharmacological effects,such as anti-anxiety,anti-obesity,anti-oxidation,anti-inflammation,enhancing immunity,protecting liver,anti-cancer and so on.Studies in recent years have shown that Ziziphus jujuba Mill.is rich in polysaccharides,flavonoids,phenolic acids,alkaloids,cyclic nucleotides,triterpenoids,vitamins and minerals and other nutrients.Most of the studies pay close attention to polysaccharides,flavonoids,phenolic acids,cyclic nucleotides and other bioactive components of jujube at home and abroad,while a few studies on triterpenoids in jube have been reported abroad.In view of the different origins and types of jujube in China,the composition,content,efficacy and mechanism of triterpenoids is different.Therefore,the study of triterpenoids from major jujube varieties in China is very important for the development of high-end functional foods and medicines of jujube.Therefore,in this study,the extraction and purification technology of triterpenoids from Laoling Jinsi jujube(Also known as Laoling Jinsixiaozao,Laoling golden silk jujube)was researched.The chemical composition analysis and identification of triterpenoids from Ziziphus jujuba Mill.cv.Jisixiaozao(ZTF)were carried out.Furthermore,the activity and mechanism of triterpenoids on prostate cancer cells were further studied.Study on Extraction and purification of triterpenoids from Laoling Jinsi jujube includes comparison of extraction and purification processes,investigation of extraction and purification pro cess parameters to determine more suitable extraction and purification conditions.We have selected the ultrasonic-assisted ethanol extraction method from various kinds of extraction process for jujube triterpenoids,and investigated the effects of key parameters such as solvent,time and material-liquid ratio on the yield of triterpenoids in jujube.Use a Ikali-soluble acid precipitation and extraction as the purification technology.Ultraviolet spectrophotometry,reversed-phase high performance liquid chromatography(HPLC-DAD)and liquid chromatography-mass spectrometry(LC-MS/MS)were used for the analysis and identification of ZTF.The new HPLC-DAD and LC-MS/MS methods can simultaneously analyze and identify seven triterpenoids in ZTF.Before the content analysis of triterpenoids in Ziziphus jujuba,the HPLC methodology was validated.Within the linear range of standard curve of each target compound,the linear correlation coefficients were above 0.999.The RSD values of intra-day and inter-day precision were 0.06?1.7%and 0.14?1.5%,respectively.The RSD values of repeatability and stability were less than 1.87%.and the recoveries of standard addition were 95.8%?102.6%.The results showed that the method is rapid,accurate and reproducible.Quantitative analysis showed that the contents of ursolic acid(202.4 mg/g)and oleanolic acid(153.4 mg/g)were the highest in the triterpene extracts of Ziziphus jujube.The contents of oleanolic acid(59.7 rmg/g)and ursolic acid(48.0 mg/g)were followed.Whatsmore,studies on the anti-prostate cancer activity and mechanism of ZTF include the establishment of PC-3 and LNCaP cell models of prostate cancer,the study of inhibition rate and apoptotic pathway of prostate cancer cells.The results showed that ZTF could significantly inhibit the proliferation of prostate cancer cells in a concentration-and time-dependent manner.The inhibition rate of PC-3 cells at 48 h was 91.5%,and IC50 was 21.2 ?g/mL.A similar trend was also observed in LNCaP cells,with the inhibition rate of 69.6%and IC50 of 48.3 ?g/mL at 48 h.Cell cycle studies showed that ZTFcould increase the number of PC-3 cells in G1 phase and decrease the number of cells in G2/M phase,suggesting that the cell cycle process might be blocked by triterpenoid extracts from Z.jujuba Mill.cv.Jisixiaozao.In addition,the contents of caspase 8,caspase 3 and caspase 9 increased with the concentration of ZTF,and following activated endogenous and exogenous pathways.In summary,this is a systematically report about the chemical composition and anti-prostate cancer capacity assessment of ZTF.We toke Z.jujuba Mill.cv.Jisixiaozao.as the research object,firstly explores the extraction and purification technology and key parameters of triterpenoids in Z.jujuba Mill.cv.Jisixiaozao..And then carries out qualitative and quantitative analysis and testing of triterpenoids.The data show that the types and contents of triterpenoids in Z.jujuba Mill.cv.Jisixiaozao.in China are different from those of other Ziziphus jujuba varieties.Finally,the inhibitory activity and mechanism of triterpenoid extracts on PC-3 and LNCaP cells were studied.The results showed that triterpenoids of Chinese Z.jujuba Mill.cv.Jisixiaozao.had a strong inhibitory effect on prostate cancer cells,and could inhibit the proliferation ofprostate cancer cells by activating two apoptotic pathways,endogenous and exogenous.Therefore,the preliminary results of this study reveal that the triterpene extract of Ziziphus jujuba has the potential to become a new therapeutic drug for prostate cancer,and the results of this study provide experimental techniques and methodologies with high practical value for further development of triterpenoids of Ziziphus jujuba. |
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