Structure Characterazation And Anticoagulant Effect Of Polysaccharides From Rosae Chinensis Flos

Rosa Chinensis Flos,an edible flower of Rosa chinensis Jacq.(Rosaceae),which was included in the 2015 edition of "Chinese Pharmacopoeia".It has the effect of promoting blood circulation and regulating menstruation,soothing liver and relieving depression.Rosae Chinensis Flos was mainly used to treat qi stagnation and blood stasis,irregular menstruation,dysmenorrhea,amenorrhea and chest pain.About its chemical composition,at present,research was on the small molecule chemical components,including flavonoids,pentacyclic triterpenes and phenolic acids.However,there were few studies on macromolecular components such as polysaccharides.Therefore,this paper selects Rosae Chinensis Flos as the research object to carry out the research on the polysaccharide content and biological activity contained in it.First of all,the crude polysaccharides from Rosae Chinensis Flos was prepared by water extraction and alcohol precipitation,petroleum ether defatting,and ethanol removal of small molecules.The crude polysaccharides were further separated and purified by DEAE-52 anion exchange column,DEAE-Sepharose FF chromatography column and Sephadex G-100 chromatography column.Two polysaccharides,RCJW-IA(molecular weight 1.2 k Da)and RCJ2-Ib(molecular weight 3.3 k Da)were obtained.Acid hydrolysismeth,ylation analysis,GC-MS,GC,FT-IR,and NMR were used to explore the structural characterization,which showed that RCJ2-Ib had a partial structure?4)-?-D-Manp-(1?linked mannan uronic acid,the monosaccharide composition of RCJW-IA was arabinose,fructose,xylose,mannose,glucose and galactose,with a molar ratio of 0.49:0.17:2.51:7.85:4.08:2.07.Secondly,the rat's acute blood stasis model was established to investigate the anticoagulant activity and mechanism of RCJ2-Ib.The results showed that RCJ2-Ib had good antithrombotic activity in vivo mainly through the following three pathways.First,it mainly acted through endogenous,exogenousand coagulation common pathway to achieve the effect of antithrombotic.The second,the formation of thrombus was inhibited by reducing TXB2 and ET-1 in serum,increasing the content of e NOS and 6-Keto-PGF1 a,as well as maintaining the balance of 6-keto-PGF1a/TXB2.The third,the anticoagulantmechanism of RCJ2-Ib may decrease WBV,HCT,PV and ESR.Last,the Illumina Mi Seq/Nova Seq designed PE250 high-throughput sequencing platform was used to sequence the V3 and V4 regions of the 16 S r DNA of mouse intestinal microorganisms.After optimizing the sequences by 97% similarity OTU classification,a total of 1927 OTUs was obtained.According to the alpha diversity index and PCo A analysis,it was found that the medium-dose group(100mg/kg)of RCJ2-Ib could maintain the diversity,stability and richness of intestinal flora,and the dose group(100 mg/kg)was more beneficial to maintain the abundance of intestinal flora in normal rats at the level of phylum and genera.