Ex Vivo To In Vivo Extrapolation Of Ferulic Acid,the Protagonist Of Polyphenolic Acid Compound:mRNA-LncRNA-kinase Regulatory Network

Background:Based on the BAP strategy,we have determined the effective components of GXII/grape juice absorbed into the blood through UPLC-QDA.After pharmacodynamic comparison,we have determined the representative components of GXII and grape juice for vasodilation,FTA and SF,but their specific relaxation The vascular mechanism is unclear.At the same time,we found that although the bioavailability of ferulic acid is low,it exerts most of the curative effect of the mother formula.In the previous stage,the research team also confirmed the effect of ferulic acid in anti-oxidation and vasodilation through experiments on VTI in vivo,in vitro blood vessels and endothelial cells,as well as the effect of ferulic acid through Ghrelin-Src/PI3k/Akt/eNOS phosphorylation pathway.Acid vasodilation mechanism.Aim:Based on previous experiments,I will further study the vasodilation mechanism of FTA/SF,a representative component of GXII/grape juice,and understand the role of ferulic acid in the regulation of oxidative stress endothelial cell model under the condition of perfecting the grape juice methodology..At the same time,it plans to discover new genes/proteins through transcriptome and phosphorylation proteomics,and improve the existing molecular signaling pathways.Methods:1.Determine the precision,stability,recovery rate and matrix effect of the 6 components of grape juice within days and days.2.Simultaneously verify the GPSDS/CPSDS and its main absorption component FTA/SF on isolated blood vessels and endothelial cells.Vasodilation effect,and inhibit it by PP2,Wortmannin,L-NAME,and explore its vasodilation mechanism.3.Establish a model of the effects of ferulic acid on healthy/oxidative damage to endothelial cells,observe the dose/time dependence of ferulic acid and the oxidative damage model to establish the relationship between its H2O2 concentration and time.4.FA-mediated oxidative damage of endothelial cells LncRNA transcriptomics and phosphorylation proteomics research.Results:1.The methodology of the six components of grape juice has suitable precision,linear range,stability,extraction rate,recovery rate,and no obvious matrix component interference,each precision is less than 15%;2.GPSDS/CPSDS and Its main absorption component,FTA/SF,can significantly relax the isolated vascular ring after PE pre-constriction.At the same time,it can increase the concentration of NO and eNOS and reduce the content of ROS for oxidative damage to endothelial cells.In the pretreatment of PP2,Wortmannin,L-NA inhibition After administration,its various effects are antagonized;3.Ferulic acid can treat H2O2-induced oxidative stress model,increase NO concentration,reduce ROS content,and is time-dose-dependent,but it is not effective for healthy endothelium;The short-term(1h)therapeutic effect and the long-term(12h)protective effect are better;the eNOS inhibitor L-NAME can significantly inhibit the eNOS expression of ferulic acid on oxidatively damaged endothelial cells;4.RNA-seq found common anti-mRNA 1170 There are 426 common anti-lncRNAs.After enriching the above genes through the KEGG pathway,we found Necroptosis(necrosis signaling pathway),mTOR(mammalian rapamycin target signaling pathway),and MAPK(mitogen activated protein kinase)signaling Pathway,cAMP(cyclic adenosine 3’,5’-monophosphate)signaling pathway,etc.5.Proteomics identified 20 differentially phosphorylated proteins as common genes for ferulic acid to regulate endothelial function.Among them,there are 11 differentially modified proteins that rise first and then fall,and 6 differentially modified proteins that fall first and then rise.KEGG The main pathways for enrichment are mTOR signaling pathway,sphingolipid metabolism,amino sugar and nucleotide sugar metabolism,and phospholipase D signaling pathway.At the same time,proteomics also found that APRT,Dnmtl,Rps3,Sept2,Rifl,Itrp3,Pxn and other common anti-proteins are related to oxidative stress,which are important phosphorylated differential proteins that need attention in the next step.Conclusion:This experiment has perfected the methodology of the 6 components of grape juice,determined the vasodilation mechanism of Guanxin-2/Grape juice’s main absorption component FTA/SF,and established a mature endothelial cell oxidative damage cell model system,and confirmed Asafoetida The protective effect of acid treatment lays the foundation for later experiments.At the same time,based on mRNA and LncRNA transcriptomics,phosphorylation proteomics screened ferulic acid to regulate endothelial cell oxidative damage related differential genes,and through bioinformatics to understand the general function of ferulic acid-mediated oxidative damage of endothelial cells.