| Propolis is a mixture formed by Apis mellifera collecting plant resin and mixing the secretions of glands such as the palate and wax glands.Propolis is a popular folk medicine and has a long history of application,which has a wide range of biological activities and rich in chemical components.The composition of different types propolis is very different,and there are many factors can affect the chemical composition of propolis,such as plant sources,collection time,geographic location,and bee species.Among them,the plant source is the most important factor.Populus type propolis are the most widely distributed over the world,and Chinese propolis typically classified as this type.But China has a vast territory and abundant plant resources.We first compared the compounds between Chinese propolis,Brazilian green propolis and Australian propolis,and used similarity analysis and cluster analysis to analyze the differences in the fingerprints of propolis in different provinces in China.The contents of p-coumaric acid and CBE were found in Changbai Mountain propolis were richer,and the CBE compound was successfully isolated,purified and prepared.The compound was identified as benzyl p-coumaric acid by mass spectrometry(MS)and nuclear magnetic resonance(NMR).In addition,the inhibitory effect of Changbai Mountain Propolis(CBMP)on seven different cancer cell lines was demonstrated,found that CBMP could significantly inhibit the proliferation of SGC-7901 cells.The main research results are as follows:1.Differences in chemical composition of Chinese propolis,Brazilian green propolis and Australian propolisThe analysis of Chinese propolis,Brazilian green propolis and Australian propolis using HPTLC and HPLC showed that the chemical composition was quite different.The chemical composition of different Brazilian green propolis samples was basically the same,the main compounds are phenolic compounds:atipicillin C,p-coumaric acid,isochlorogenic acid A and isochlorogenic acid C;Chinese propolis were consistent with the characteristics of poplar-type propolis,but the contents of different compounds are different,the main components are phenolic acids and flavonoids;Australian propolis shows a large difference between different samples,two samples are poplar type propolis and others are unknown types.2.The chemical composition,antioxidant activity and spectrum-effect relationship of Chinese propolisAll 49 propolis samples from different regions of China were tested by HPLC-UV and were consistent with the characteristics of poplar-type propolis.Similarity analysis and cluster analysis revealed that some samples collected from Northeast China were significantly different from other samples.The DPPH was used to test the antioxidant capacity of 49 Chinese propolis,and the used principal component analysis(PCA)and multiple linear regression analysis(MLRA)to obtain the spectrum-effect relationship between the chemical compositions and antioxidant activities of Chinese propolis;establish the off-line DPPH-HPLC assay,by comparing the changes of HPLC chromatographic peaks in samples before and after the reaction with DPPH solution.Eight compounds(caffeic acid,ferulic acid,kaempferol,unknown compound 1,benzyl caffeate,pinobanksin 3-oacetate,CAPE and galangin)showed a decrease in peak area after the reaction,indicating that these compounds have antioxidant activity.3.The chemical composition and plant sources of CBMPThe chemical composition analysis of 104 propolis samples collected from Northeast China,demonstrated that CBMP were significantly different from propolis in other regions.The contents of p-coumaric acid and CBE compounds in CBMP were highe.HPLC analysis showed the botanical sources of the CBMP are P.davidiana dode and P.simonii Carr.4.Separation,purification,preparation and chemical synthesis of compound CBEThe compound CBE was separated and purified and the high purity CBE was obtained by preparative liquid chromatography.The compound CBE was identified by HPLC-MS and NMR as benzyl p-coumarate.Benzyl p-coumarate was synthesized from p-coumaric acid by acid chloride method.5.The anti-cancer activity of CBMPDifferent cancer cell lines were treated with the ethanol extracts of CBMP for 24 hours and the cell viability was examined by CCK-8 assay.The results showed CBMP had weak activities against human pancreatic cancer cell PANC1,human lung cancer cell A549,human colon cancer cell HCT116,human liver cancer cell HepG2,human bladder cancer cell T24 and human breast cancer cell MDA-MB-231;but it significantly inhibited the growth of human gastric cancer SGC-7901 cells,caused cell apoptosis and cell cycle arrest in S phase,with increased production of reactive oxygen species(ROS)and reduced mitochondrial membrane potential(MMP).The results indicate that Chinese propolis sourced from Changbai Mountains selectively inhibits the proliferation of SGC-7901 cells by inducing both death receptor-induced and mitochondria-mediated apoptosis,and S phase cell cycle arrest. |
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