Comprehensive Analysis Of Bioactive Compounds In Xinjiang Jujube And Its Regulation Mechanism On Oxidative Damage Of Cells Induced By Benzopyrene

The southern region of Xinjiang has the geographical and climatic conditions suitable for the growth of jujube.Xinjiang jujube is rich in triterpene acids,flavonoids,cyclic nucleotides and other functional factors,which have physiological functions of antioxidant stress,anti-cancer,anti-inflammatory,immune regulation,and gastrointestinal protection,etc.Benzopyrene?Ba P?widely exists in air,water,soil,food and packaging materials,which has strong carcinogenicity,mutagenicity and indirect teratogenicity.Ba P enters the human body through food,water,respiratory system and other ways,and stimulates the body to produce a large number of free radicals,which can result in cellular oxidative stress.Oxidative stress,which is the main risk factors of human aging,tumor,cardiovascular and cerebrovascular diseases,neurodegenerative diseases and diabetes,can induce gene mutation,protein denaturation,mitochondrial dysfunction and lipid peroxidation.Therefore,to study the functional factors of jujube against Ba P harm and further exploring the molecular mechanism of its regulation of Ba P-induced oxidative damage have scientific guiding effects on the creation of new functional food with jujube as raw material.The main research contents and results of this paper are as follows:?1?The identification and multivariate statistical analysis of functional factors in Xinjiang jujube.The main functional factors?flavonoids,triterpene acids,nucleotides and adenosine compounds?in jujube were studied qualitatively and quantitatively by high performance liquid chromatography?HPLC?and mass spectrometry?MS?.The results showed that the total content of triterpene acid in jujube ranged from 1082.775±63.431–7915.451±510.824?C41,jing 39?.Betulinic acid,citric acid,cratonic acid,oleanolic acid,ursolic acid,betulonic acid and 2-hydroxyursolic acid are the main triterpene acids in xinjiang jujube.The total nucleic acid content ranged from 408.329±8.611–1030.450±24.634?g/g DW?C45,junzaobianzhong 2?.Nucleic acids mainly include uracil,xanthine,hypoxanthine,guanine and adenine.The flavonoids content ranged from 102.886±3.581–571.678±18.653?g/g DW?C23,guanyinzao?.The main flavonoids are catechin,epicatechin,p-coumaric acid,rutin,chastein,isopressin,ferulic acid,quercetin 3-O-glucoside,dihydroquercetin,naringin,quercetin,dihydropanaphthol,myriceflavone,saccharol and alicylic acid.?2?The screening of functional factors of jujube based on cell oxidation damage modelFirstly,the oxidative damage model of human lung cells induced by Ba P was established.Furthermore,the functional factors with the best regulatory effect on cell oxidation damage in jujube were screened and their intervention methods were proposed based on the model.The results showed that the cell oxidation damage model was established under the condition of Ba P as the induction reagent,the induction concentration was 400?mol/L,and the induction time was 48 h.Compared with other treatment groups,ursolic acid?UA?intervention significantly increased the proliferation rate and antioxidant enzyme activity?SOD,CAT,GSH?in damaged model cells,and decreased the contents of malondialdehyde and reactive oxygen species in cells?P<0.05?.Under the principle of choosing a relatively high concentration level with little impact on normal cell growth,UA was selected as the optimal functional factor,the optimal intervention concentration was 10?g/m L,and the optimal intervention time was 48 h.?3?Study on the regulatory mechanism of ursolic acid on cell oxidative damage induced by Ba POn the basis of the above experiments,UA was sellected as the best intervention reagent,and the effects of UA on cell cycle and apoptosis,DNA damage,autophagy related protein expression were studied,and laser scanning confocal microscope and transmission electron microscopy test were also used to explore the regulation mechanism.Compared with the oxidative damage group,the G₀/G₁phase cells in the UA intervention group increased by12.770%,the S phase decreased by 13.162%,the G₂/M phase decreased by 49.864%?P<0.01?,the apoptosis rate decreased by 58.170%,and the content of 8-hydroxydeoxyguanosine?8-OHDG?decreased by 46.39%.The results indicated that UA could significantly improve the blocking effect of Ba P on cell cycle,reduce the apoptosis rate and repair the DNA damage of cells.In addition,after ursolic acid intervention,the expression of autophagy related proteins?LC3-?/LC3-?,Beclin1,Atg5,P62?were 0.853,0.368,0.430 and 0.298,respectively,which were significantly different with the damage model group?P<0.05?.these result indicateded that UA intervention can significantly reduce the cell excessive autophagy,thus effectively protect cells from injury of Ba P.Laser scanning confocal microscopy and transmission electron microscopy further confirmed that UA can significantly reduce the oxidative damage induced by Ba P,and the mechanism is related to the inhibition of excessive autophagy.?4?Transcriptome analysis of the regulation effect of UA on the oxidative damage induced by Ba P.Transcriptome sequencing technology and bioinformatics analysis were used to study the regulatory effect of UA on the oxidative damage induced by Ba P and its molecular mechanism.The results showed that significantly up-regulated and down-regulated genes were 296 and 688,respectively.The biological processes of GO entries enriched by differentially expressed genes are mainly as follows:cell connection,intracellular receptor signaling pathway,and the regulation of i-kappab kinase/NF-kappa B.The main cell components were:nucleus,nucleus,cell membrane and cytoplasm.The main molecular functions were:insulin-like growth factor-activated receptor activity,protein tyrosine/threonine phosphatase activity,transcription factor binding.The KEGG pathway analysis of differentially expressed genes mainly enriched to:cancer signaling pathway,PI3K-Akt signaling pathway,MAPK signaling pathway,and apoptosis signaling pathway.The key differentially expressed genes were verified by q RT-PCR,the results show that the regulatory mechanism of UA on Ba P induced cell oxidative damage mainly by alleviating excessive autophagy and regulating the cellular aging process,and atg2a,nfkb1,eif4ebp1,pik3ip1 and nr4a1 genes on the PI3K-Akt/m TOR signaling pathway are mainly involved in the regulatory process.In conclusion,this study analyzed the main functional factors in different varieties of jujubes.Ursolic acid was selected as the best functional factor by the oxidative damage model induced by Ba P.The protective effect of UA on oxidative damage induced by Ba P and its molecular mechanism were preliminarily elucidated.The results provided a theoretical basis for the deep processing and utilization of jujube in southern Xinjiang,which is of great significance for the sustainable and healthy development of jujube industry in southern Xinjiang.