Based On Network Pharmacology To Explore The Material Basis And Mechanisms Of The Anti-Inflammatory And Analgesic Effects Of Paederia Scandens (Lour.) Merr.

Paederia scandens(Lour.)Merr,It is a whole herb of the Rubiaceae family and the genus Paederia scandens.It has a sweet and slightly bitter taste,and has various pharmacological effects such as sedation,analgesia,anticonvulsant activity,and antibacterial activity.Although there have been many reports on the chemical components and pharmacological effects of Paederia scandens,there is still a lack of research on the substance basis and mechanism of its anti-inflammatory and analgesic effects,greatly limiting its use in veterinary clinical practice.Therefore,based on the optimization of the preparation process of total iridoid terpenes from Paederia scandens,this experiment analyzed its chemical composition and conducted network pharmacology research.Then,molecular docking methods were used to speculate on the main active ingredients,and in vivo validation was carried out to further evaluate the impact of the active ingredients on the main signal pathways,reveal the molecular mechanisms of the anti-inflammatory and analgesic effects of the active ingredients,and lay a theoretical foundation for the clinical application of Paederia scandens in anti-inflammatory and analgesic aspects.The specific research content is as follows:(1)Optimization of the extraction process of total iridoid glycosides from Paederia scandens.Based on the single-factor experiment,the extraction rate of total iridoid terpenes from Paederia scandens was used as the evaluation index.The response surface methodology of Box Benken design with three water levels and four variables was used to optimize the extraction process of total iridoid terpenes from Paederia scandens.The optimal extraction conditions were obtained as follows: temperature 96 °C,ethanol concentration 56%,material liquid ratio of 1:26,extraction time 1.56 h,extraction times 3 times.Under this optimal condition,the experiment was repeated three times,and the actual extraction rate was 21.83±0.41 mg/g,which was close to the predicted value of 21.947 mg/g.(2)Optimization of the enrichment process for total iridoid terpenoids in Paederia scandens.On the basis of the optimal extraction process,the enrichment process of total iridoid terpenoids in Paederia scandens was optimized using the macroporous resin method.Ten types of macroporous resins were selected to investigate their adsorption characteristics,and the static adsorption results showed that SP825 macroporous resin was the most suitable.Based on the dynamic adsorption results,the optimal enrichment conditions were further optimized.The optimized adsorption parameters were as follows: The initial concentration of total iridoid solution was 2.5 mg/m L,the loading amount was 2.91 BV,and the flow rate was 2 BV/h.The optimized desorption parameters are 2 BV of deionized water,5 BV of 70% ethanol,and a flow rate of 2 BV/h.According to the optimal conditions,the concentration of total iridoid terpenes in Paederia scandens was increased by 3.7 times,with a recovery rate of 80.98%.The results showed that SP825 had a good ability to enrich total iridoid terpenes in Paederia scandens,providing a theoretical basis for the preparation of total iridoid terpenes in Paederia scandens.(3)Anti-inflammatory and analgesic activities of the extract from Paederia scandens before and after enrichment.Using xylene-induced mouse ear swelling and carrageenan foot swelling inflammation models,hot plate,acetic acid writhing,and formalin biphasic pain experiments,the anti-inflammatory and analgesic activities of Paederia scandens before and after enrichment were studied,and it was confirmed that the anti-inflammatory and analgesic effects of Paederia scandens extract after enrichment were indeed better than those before enrichment,and were dose dependent.(4)Exploring the mechanism of action using liquid chromatography-mass spectrometry combined with network pharmacology.UPLC-ESI-MS was used to identify the chemical components in the enriched extract of Paederia scandens,and a total of 18 chemical components were identified.Using network pharmacology methods to analyze 18 chemical components and inflammatory and pain disease targets.The results showed that the core targets of the anti-inflammatory and analgesic effects of Paederia scandens were SYK,TNF,COX-2,and IL-6,respectively.The main related pathway enriched by KEGG was the C-type lectin receptor signal pathway.Using molecular docking technology to analyze the binding effects of these 18 possible active ingredients with core targets,the results showed that kaempferic acid has the best binding effect,and it is speculated that kaempferic acid is the leading chemical component that plays an anti-inflammatory and analgesic role.(5)Component separation and validation of prediction results.The anti-inflammatory and analgesic activities and mechanisms of kaempferic acid were verified using the above activity experiments,q RT-PCR,and Western blot methods.The results showed that kaempferic acid has good anti-inflammatory and analgesic activities,and plays an anti-inflammatory and analgesic role by inhibiting the expression of key genes in the pathway,DECTIN-1,SYK,P65,and COX-2.The results are consistent with the network pharmacological prediction.This study not only identified the main anti-inflammatory and analgesic components of Paederia scandens,but also explored their therapeutic mechanisms,providing a prerequisite for the study of the pharmacokinetics and pharmacodynamics of the effective components of Paederia scandens in vivo,laying a foundation for the establishment of quality control methods for Paederia scandens,and making a meaningful exploration for the modernization of traditional Chinese medicine.