Mechanism Of Raphanus Sativus L.in The Intervention On High-normal Blood Pressure With Excessive Phlegm-dampness Syndrome

Objective:The purpose of this study was to explore the mechanism of Raphanus sativus L.（RS）intervention on high-normal blood pressure model rats with excessive phlegm-dampness syndrome.Methods:1.Prepared the high-normal blood pressure model rats with excessive phlegm-dampness syndrome by feeding with high-fat diet and intraperitoneal injection of Nω-Nitro-L-arginine,and evaluated the model from the indicators of blood pressure,body mass,macroscopic signs and blood biochemical indexes.2.Pharmacodynamic evaluation of drug RS on the model rats:The effect of the drug was evaluated from different levels in vivo and in vitro,such as macroscopic signs,blood pressure,pathological changes of rat thoracic aorta,blood biochemistry,endothelin-1 and nitric oxide,and measurement of vascular tension of isolated thoracic aorta.3.Integrated analysis of high-throughput transcriptomics and metabolomics:The differentially expressed genes（DGEs）and differential metabolites after RS intervention model and the relationship between them were analyzed,so as the molecular mechanism of RS intervention on high-normal blood pressure model rats with excessive phlegm-dampness syndrome were reveal.Results:1.The model rats had dark hair color,lazy movement,poor appetite and loose stool.The body mass,blood lipid of model rats was significantly increased,at the same time the blood pressure is slightly increased and maintained stable at about 160/88mm Hg for seven weeks.Therefore,this method is feasible to construct the rat model of high-normal blood pressure with excessive abundant phlegm-dampness syndrome.2.The blood pressure of model rats decreased significantly,the blood lipid content and damaged vascular endothelium were improved,while endothelin-1 decreased significantly and nitric oxide increased significantly after the intervention of RS.The vascular tension test of thoracic aorta in vitro further confirmed that RS could reduce blood pressure through improving endothelial dysfunction.3.Using the RNA-seq technology and UPLC-MS/MS metabolomics analysis,a total of 65 DGEs(P_adj<0.05 and fold change（FC）>2)and 52 differential metabolites（VIP>1,P value<0.05 and FC≥2）were found to be remarkably altered by RS intervention,in addition,fully connected network between DGEs and metabolites was investigated.It was found that RS may prevent vascular endothelial dysfunction to antihypertension by regulating the signaling pathways including purine metabolism,linoleic acid metabolism,arachidonic acid metabolism,circadian rhythm and phosphatidylinositol and Pik3c2a,Hspa8,Dnaja1,Arntl,Ugt1a1,Dbp,Rasd1,Aldh1a3 were found to be the putative target genes.Conclusions:RS could mitigate the vascular endothelial dysfunction of high-normal blood pressure model rats with excessive phlegm-dampness syndrome by regulating purine metabolism,linoleic acid metabolism,arachidonic acid metabolism,circadian rhythm and phosphatidylinositol signaling pathways and Pik3c2a,hspa8,Dna JA1,Arntl,Ugt1a1,Dbp,Rasd1 and Aldh1a3 may be the putative target genes.