Optimization Of Extraction Conditions Of Mogroside Ⅴ And Its Bioactivity Analysis

Siraitia grosvenorii is an important plant resource of medicinal and edible in China.Mogroside V（MV）contained in it,a natural sweetener,and has been widely used in modern food processing.Meanwhile,various pharmacological activities and functional effects.However,the effect of MV have not been relatively analyzed and studied on the genome-wide expression of human hepatocytes.In this study,MV was used as the research object for experimental investigation.Firstly,to explore the effect of different extraction factors on the extraction rate of MV,and then to evaluate the antioxidant activity of five purity MV,finally the genome-wide analyzed based on transcriptome sequencing technology,and targeting explore the effects of MV on DEGs and typical signaling pathways related to inflammation,antioxidant and glycolipid metabolism.The purpose is further understanding the biological activity of MV on human hepatocytes,and providing a theoretical and reference basis for further development and research in the food-medicine field and functional food resources.Furthermore,providing a new idea accelerate the comprehensive development of activities composition in Siraitia grosvenorii.The main results were as follows:1.Optimizing extraction process and condition about Mogroside V.Explored the effects of the four single factors of juice extraction rate,solid-liquid ratio,extraction time and temperature on the extraction rate of MV.On this basis,these were carried out to further optimize by orthogonal experiments.The study results show that juice extraction rate is the most significant factor of influence,followed by the material-liquid ratio,extraction temperature,and the extraction time has a relatively small effect.By optimizing,the best extraction process is juice extraction rate of 25%,solid-liquid ratio of 1:5,extraction temperature of 80 ℃,and extraction time of 90 min.Extraction rate up to 97.43%.2.Evaluation and analysis of antioxidant activity about Mogroside V with five kinds of purity.The five kinds of purity（20%,30%,50%,60%,95%）MV were respectively investigated with five kinds（DPPH,ABTS,FRAP,ORAC and superoxide anion）antioxidant test methods.The results of the five methods show that,under the same test conditions,the greater the purity of MV has the weaker the antioxidant activity.20% and 95% are nearly 13 times different in ORAC.The antioxidant activity of MV with different purity is in order: 20%>30%> 50%> 60%> 95%.The above results indicate that the MV in the extract has weak antioxidant activity in vitro,and there may be other active substances or synergistic antioxidant components,which exert more significant antioxidant capacity.3.Using RNA-seq to study the effect of MV on the genome-wide expression of Hep G2 cells,and in-depth analyze its antioxidant and anti-inflammatory function in the cellular and gene level.Combined with bioinformatics analyze DEGs,obtained 691 genes with significant changes,accounted for 1.52% of the total genes（p <0.05）.Among them,there are 39 upregulated and 53 down-regulated genes that have more than two changes,respectively accounting for 0.56% and 0.96%;By using GO functional enrichment analysis,Multiple subfunctional rich regions are related to formation and transmission of RNA,to translation and transport of protein,moreover,the biological processes（BP）have most types involving up to393;The metabolic pathways of KEGG enrichment analysis found that 7 pathways have extremely significant difference changes.Among them,RNA transport is the most significant signal pathway involving 20 DEGs,and it has also been found to involve multiple human disease metabolic pathways.Among the DEGs,3 are related to antioxidation,and DHRS1（115817）is the most up-regulated gene by 16 times;other 8 are also related to inflammation,and TRIM27（5987）is the most down-regulated by 57 times.We found that most of the effects of MV on processes of cellular metabolism are achieved by down-regulating gene expression,besides these genes are less affected related to oxidation,and most are related to inflammation.4.The effect that Mogroside V on the expression of genes is related to glycolipid metabolism in Hep G2 cells.We collect 4 up-regulated and 6 down-regulated genes.Among them BTC（685）was the up-regulated gene with the largest fold change,up to 26 times,besides CFHR1（3078）was the down-regulated gene with the largest,up to 6 times;Similarly by the KEGG analysis,it was found that non-alcoholic fatty liver disease（NAFLD）had extremely significant changes,involving 15 DEGs,and followed by glycosphingolipid biosynthesis-lacto and neolacto series.The results indicate that MV can regulate glycolipid metabolism.