Effect Of Gardenia Crocetin On The Whole Genome Expression Profile Of Human Hepatocellular Carcinoma Cells And Its Application In Dairy Products

Gardenia fruit extract is rich in crocin,crocetin and its derivatives,and the mainly active ingredients include crocetin.Crocetin,a type of long-conjugated water-soluble carotenoid,was early derived from the stigma of saffron and the fruit of gardenia,which is used in traditional Chinese medicine to protect the cardiovascular,anti-tumor,lower blood sugar,and inhibit cholesterol synthesis.However,the function of crocetin on human hepatocytes is rarely studied due to the preciousness of saffron medicinal materials and lower yields at present.In this study,we performed transcriptome sequencing(RNA-seq)analysis of gardenia extract crocetin before and after treatment of Hep G2 cells.We identified differentially expressed genes(DEGs)related to the effect of crocetin on Hep G2 cells and then performed function annotation and pathway analysis of DEGs.Based on the results of these bioinformatics analysis,predicted the biological functions of the differential genes and selected the core genes and key genes related to crocetin treatment,and that performed the specific mechanism analysis.Finally,the characteristics of functional yoghurt added with crude extract of gardenia fruit rich in crocetin were studied.Our study aims to explore the influence of crocetin on the whole genome expression of human hepatocytes and the physiological function of the body through nutritional genomics,molecular biology,bioinformatics and related analysis tools,and also provide a theoretical basis for the development of related fermented milk products.(1)Crocetin can affect the expression of 19,821 genes in 22580(annotated genes)in human hepatocellular carcinoma cells.By analyzing the alternative splicing pattern of the gene transcripts through r MATs software,it was found that 9,634 genes have alternative splicing.Exon skip types accounted for the largest proportion,nearly 65%,and intron retention types accounted for the least proportion,nearly 3%,The main enrichment pathways of differential alternative splicing genes include transcriptional regulation and autophagy regulation(2)Using DESeq2 for differential analysis,774 differential genes were identified,of which 491 genes were up-regulated and 283 genes were down-regulated.GO functional analysis and KEGG pathway enrichment analysis These differential genes are mainly concentrated in cell proliferation and apoptosis(such as BRD4 and DAXX),lipid formation(such as EHMT2),cell response to growth factor stimulation(such as CYP24A1 and GCNT2),and growth Factor binding(such as ABCB1 and ABCG1),metabolism and signal transduction processes.(3)After constructing a PPI network using STRING online protein interaction database,draw a PPI network map using Cytoscape software,and obtain 21 core proteins from high to low according to the degree of node degree.Core proteins are mapped to modules,and it can be seen that most core proteins are in the above modules.Real-time fluorescence quantitative analysis confirmed that the transcriptome sequencing data was consistent with the experimentally verified data.(4)To explore the effect of adding gardenia fruit crude extract(SGE)which is rich in crocetin on the fermentation quality of yoghurt.We found that SGE can effectively promote proliferation of Bulgarian bacilli in yofurt and maintain the stability of the strain in the initial storage period,and also promote the formation of the characteristic flavor substances aldehydes and ketones in yogurt and provide the product a better egg yellow color.In addition,the addition of SGE has obvious enhancement effects on the bacteriostasis,gastrointestinal fluid tolerance and antioxidant activity of yogurt system.Therefore,proper addition of SGE can not only significantly improve the quality of yogurt,but also enhance the nutritional value of yogurt.Conclusion: Gardenia fruit extract crocetin mainly affects the physiological and metabolic processes of human hepatocytes through the following processes,most of which are positive promotion effects,such as: pro-inflammatory factor expression,drug metabolic enzyme synthesis,glycolipid metabolism,cell proliferation And apoptosis.And by adding gardenia fruit extract(rich in crocetin)to yoghurt,it was found that it can enhance the fermentation process of yoghurt and provide a molecular research basis for the subsequent advancement of saffron acid dairy products.