Mechanism Of Cucurbita Ficifolia Bouché On Prevention And Treatment Of Obesity Based On Network Pharmacology And Molecular Docking Technology

Cucurbita ficifolia Bouche has a variety of biological activities and has broad development prospects.In this paper,Cucurbita ficifolia Bouche was used as a raw material to assist the extraction of flavonoids from Cucurbita ficifolia Bouche by ultrasonic method,and the extraction process was optimized using response surface methodology Ultrasonic assisted extraction of flavonoids from Cucurbita ficifolia Bouche,using response surface method to optimize the extraction process Separation and purification of flavonoids from Cucurbita ficifolia Bouche by macroporous adsorption resin.HPLC-MS/MS technique was used to analyze its components.Based on network pharmacology and molecular docking technology,the weight loss mechanism of flavonoids in Cucurbita ficifolia Bouche was analyzed.The biological activities of flavonoids from Cucurbita ficifolia Bouche was investigated.The results of the study are as follows:The influence factors were determined by single factor experiment,and the extraction process were optimized by response surface methodology:ultrasonic power 250 W,ethanol volume fraction 70%,ultrasonic time 50 min,solid-liquid ratio 1:20 g/m L,and the extraction yield of flavonoids from Cucurbita ficifolia Bouche reached 3.531%.AB-8macroporous resin was used to purify flavonoids from Cucurbita ficifolia Bouche.The technological conditions were:The extraction rate of flavonoids was 4.517～4.609%under the conditions of loading flow rate 1.0 m L/min,loading solution p H3.0,elution solution80%ethanol,elution flow rate 1.0 m L/min.By HPLC-MS/MS analysis of its composition,a total of 6 flavonoids were obtained,including quercetin,rutin,persioside,luteolin-7-O-glucoside,hyperin and methyl lostimarin,among which the content of quercetin was the highest,accounting for 45%.The active components,potential targets and molecular mechanism of flavonoids from Cucurbita ficifolia Bouche were predicted by network pharmacology method.Construction of the"component-target-pathway"interaction network between flavonoids and obesity,including drug similarity assessment,oral bioavailability prediction,protein-protein interaction（PPI）network construction and analysis,gene ontology（GO）terminology and reaction group pathway annotation.Ultimately determine the Cucurbita ficifolia Bouche flavonoids of 5 kinds of active ingredients regulate the 5 key targets of obesity related with eight major signaling pathways,GO bioaccumulation analysis results show that the Cucurbita ficifolia Bouche flavonoids weight loss mainly involved the regulation of obesity-related redox responses,inflammatory reaction,and the proliferation,cycle,differentiation of fat cells.Molecular docking simulation was used to further explain the active site and binding degree of ligand and explain its possible mechanism of action.The results showed that the core target had higher binding energy with luteolin-7-O-glucoside and persioside.The antioxidant properties of Cucurbita ficifolia Bouche flavonoids were investigated by free radical scavenging assay,and the antioxidant activity increased with the increase of flavonoid concentration.In vitro cell experiments showed that flavonoids of Cucurbita ficifolia Bouche could inhibit the secretion of nitric oxide（NO）and prolandin E₂（PGE₂）from mouse mononuclear macrophages（RAW264.7）induced by lipopolysaccharide（LPS）,proveding that flavonoids of Cucurbita ficifolia Bouche had anti-inflammatory activity,and the anti-inflammatory ability was proportional to the concentration of flavonoids of Cucurbita ficifolia Bouche.Flavonoids of Cucurbita ficifolia Bouche can effectively inhibit the proliferation and apoptosis of 3T3-L1 pre-adipocytes,and inhibit the differentiation of3T3-L1 pre-adipocytes and the production of adipocytes.The cells were treated with 50μg/m L,100μg/m L,200μg/m L,400μg/m L flavonoids of Cucurbita ficifolia Bouche,and the experiment proved that the fat content of the cells decreased by 10.57%,14.2%,19.5%,26.7%,with significant differences compared with the blank control group.