Research On The Glycosides Extraction Of Rubus Chingii Hu And Its Activity Against Ulcerative Colitis

In this study,rubus chingii Hu was used as raw material to extract glycosides from rubus chingii Hu through ultrasonic assisted extraction,response surface method was used to optimize the extraction process,macroporous adsorption resin was used to separate and purify rubus chingii Hu glycosides,and the process was optimized.Its components were analyzed by HPLC-ESI-MS.Mechanism analysis of rubus chingii Hu glycosides against ulcerative colitis based on network pharmacology and molecular docking technology;And investigate its activity.The research contents are as follows:The influencing factors for the extraction of rubus chingii Hu glycosides were determined through single factor experiments.The best processes optimized by response surface method were:liquid ratio 1:27 g/m L,ethanol concentration 57%,ultrasonic time was 28 min,ultrasonic power was 400 W,and the extraction amount of glycosides in rubus chingii Hu was 196.8 mg/100 g.The Rubus chingii glycosides were purified by HP-20macroporous resin under p H 4.0,6.5 BV,concentration of 2.0 mg/m L,loading flow rate of1.5 SV,80%and elution flow rate of 1.0 SV with extraction of 182.6（±0.51）mg/100 g.The compositions of rubus chingii Hu glycosides were analyzed by HPLC-ESI-MS.And seven rubus chingii Hu glycosides were obtained,namely hypericin,ellagic acid,Quercetin-3-O-rutinose,Miquelianin,kaempferol-3-O-rutinoside,Astragalin and Tiliroside,among which ellagside had the highest content,accounting for 47.65%of the glycosides.Network pharmacology was used to virtually screen the active components,potential targets and mechanism of action of the rubus chingii Hu glycosides,and 26 core targets and 10 major signaling pathways were selected,proving that the rubus chingii Hu glycosides can jointly regulate the anti-ulcerative colitis mechanism with"multi-component,multi-target and multi-pathway".Using molecular docking technology,the anti-ulcerative colitis mechanism of rubus chingii Hu glycosides was clarified by the degree of ligand and receptor binding,and the results showed that the core target and the compound ellagin had high binding energy.In vitro inflammation model was established to investigate the inhibitory effect of rubus chingii Hu glycosides on NO and PGE₂secreted by mouse macrophages（RAW264.7）and mouse embryonic fibroblasts（NIH3T3）induced by LPS.The results showed that rubus chingii Hu glycosides had anti-inflammatory activity.By establishing a model of ulcerative colitis in vitro,the inhibitory effect of rubus chingii Hu glycosides on interleukin-1（IL-1β）and interleukin-6（IL-6）secreted by human colorectal cancer cells（Caco-2）induced by LPS was investigated,indicating that rubus chingii Hu glycosides had anti-ulcerative colitis activity,and the inhibitory effect was proportional to the concentration of rubus chingii Hu glycosides.Rubus chingii Hu capsules were prepared from rubus chingii Hu glycosides.The excipients were added and granulated by measuring the Angle of reactivity and pile density.The results were as follows:the ratio of raw material to auxiliary material was 4:1,the ratio of auxiliary material（soluble starch:dextrin）was 1:2,and the volume fraction of ethanol was 90%.Capsule appearance is not damaged,load difference,disintegration time do not exceed the relevant regulations.