The Accumulation Of Blueberry Anthocyanin And The Effects Of Its Extracts On Colitis Associated-colorectal Cancer And Related Mechanism

Blueberry fruits taste sweet and sour,with the high commercial and nutritional values,which are directly edible and use to be made products.Berries from different geographical locations,cultivars and development stages may display substantial differences in the quality parameters.Study on how cultivars and ripening can alter taste and phytochemicals in blueberry,which can help guide the industry for optimum conditions during harvesting.Anthocyanin can directly affect the functional and nutritional effects,which display a wide range of biological activities including free radical scavenging,metabolic regulation,and health-protective effects.Inflammatory bowel diseases(IBDs)have already become one of the major gastroenterological problems.The increased degree of inflammation and duration of disease largely increases the risk of colon cancer,which extremely influence the quality of patient's life.Therefore,the study on the inhibitory mechanism of blueberry anthocyanin in colon and colitis associated colorectal cancer(CAC),which provides theoretical references for seeking a potential drug or functional product for improving gut health,is of great significance.Comparative study of the anti-inflammatory properties of BE in vitro and in vivo was conducted on HCT-116 colon cancer cells,lipopolysaccharide(LPS)-activated mouse macrophages,and on azoxymethane(AOM)/dextran sodium sulfate(DSS)-induced CAC in miceThe main results are as follows:Variations in anthocyanin and phenolic compounds content in Bluecrop and Northblue blueberry cultivar fruits were studied,and comparative transcriptome analysis was performed to analyze differences in the molecular mechanisms of anthocyanin biosynthesis.A total of 13 799 unique genes were identified by differential expression analysis,and further subjected to GO classification and pathway enrichment.Nine differentially expressed genes(DEGs),including CHI,DFR,F3?H,FLS,CHS,OMT,UGT,ANS and F3 H,were selected to validate the differential expression data using quantitative real-time PCR.The obtained q RT-PCR expression results were consistent with the RNA-Seq results.The expression levels of 9 candidate genes involved in flavonoid biosynthesis and metabolism were concurrent with the anthocyanin content.The developmental stage appeared to affect the expression of genes related to flavonoid biosynthesis to a greater extent than the tissue or cultivar type.Ripening of fruit can lead to changes in quality parameters in blueberries.The objective was to determine the effect of ripening on taste and phytochemicals of two blueberry cultivars.Blueberry fruits of Bluecrop and Northblue cultivars were divided into five ripening stages.Taste profiles,phytochemicals and antioxidant activities of blueberry fruit were evaluated.During ripening,sweetness,sourness,and astringency showed a great response.Fruit color significantly changed from green to blue-purple,with anthocyanins,proanthocyanidins,flavonoids and polyphenol compounds significantly altered.The contents of delphinidin-type and malvidin-type anthocyanins,which are closely associated with blue-purple color,increased from first to fifth stage of maturity.Changes on antioxidant activity during ripening were similar to total anthocyanins.Regardless of maturity stage,Northblue cultivar displayed higher concentrations of phytochemicals and antioxidant activity.The results demonstrated that treatments of BE/OX showed inhibitory effects on HCT?116 cell and nontoxic effect on CCD?18Co normal colon cells.Flow cytometry analysis indicated that treatment with the BE,OX or in combination could induce G0/G1 cell cycle arrest,apoptosis,increase of reactive oxygen species,and induce loss of mitochondrial membrane potential in HCT?116 cells.Furthermore,after treatments,the expression of inflammatory cytokines was decreased,cyclin D1 and CDK4 were decreased;caspases?3 and 9 were activated;the Akt/Bad/Bcl?2 pathway was modulated.Moreover,the combination treatment had a considerably higher growth inhibitory effect on human colon cancer HCT?116 cells than that of BE or oxaliplatin alone.BE reduced the production of pro-inflammatory markers in LPS-induced macrophages.On the other hand,gavage feeding administration of BE/MES in AOM/DSS induced CAC mice lead to colon shortening,elevated disease activity index and tumor loading,presence of blood in the stool,loss of weight,through blocking pro-inflammatory cytokines,the suppression of NF-?B signaling,decrease the marker of anti-apoptosis and cell proliferation.BE/MES may be of protentional utility in the chemoprevention of CAC.