Preparation, Component Characterization And Anti-pelvic Inflammatory Disease Research Of Total Phenolic Parts Of Prunella Vulgaris Stems And Leave

Objective:Prunella vulgaris L.,a perennial herb of Prunella vulgaris L.genus in Lamiaceae,is commonly used as a traditional Chinese medicine in China.The common medicinal part of Prunella vulgaris L.is dry ears,which only accounts for 20%of the whole grass,so it may cause a large waste of resources.At present,the research on Prunella vulgaris L.is mainly focused on the pharmacological action and molecular mechanism of Prunella vulgaris L.ear,and the research on non-medicinal parts is less.The preliminary laboratory research found that Prunella vulgaris L.stem and leaf total phenols have anti-inflammatory effects through pharmacological experiments.In order to explore the mechanism of Prunella vulgaris L.stem and leaf total phenols to play an anti-inflammatory role,the chemical components contained in it,the targets and pathways of action,The changes of metabolites caused by the action on the body were studied,which laid a scientific foundation for the development of Prunella vulgaris L.stem and leaf resources,and provided a reference for the study of the anti-inflammatory pharmacological mechanism of Prunella vulgaris L..Methods:1.The extraction process of Prunella vulgaris L.stem and leaf total phenols was optimized by response surface methodology,and the qualitative and quantitative analysis methods of Prunella vulgaris L.stem and leaf total phenols were established by UPLC-Q-TOF-MS/MS and HPLC.2.Through network pharmacology,Macromolecular docking and molecular dynamics simulation,the anti-inflammatory mechanism of Prunella vulgaris L.stem and leaf total phenols identified in"1"was explored,and related targets and pathways were predicted.3.Mechanical injury and bacterial infection were used to replicate the chronic pelvic inflammatory rat model to verify the efficacy of Prunella vulgaris L.stem and leaf total phenols in the treatment of chronic pelvic inflammatory disease,and Transcriptome was used to explore its mechanism from the genetic level.4.The metabonomics of the serum of rats in"3"was studied by GC-MS,in an attempt to find the anti-inflammatory mechanism of Prunella vulgaris L.from the changes of metabolites.Results:1.The best extraction process of Prunella vulgaris L.stem and leaf total phenols was:30times water(V·m^-1),extraction time 120 min,extraction 3 times,which was proved to be simple and feasible.22 compounds in total phenols of Prunella vulgaris L.stem and leaf were identified,including 16 phenolic acids and 6 flavonoids.The content of caffeic acid and rosmarinic acid was2.33 mg·g^-1 and 7.37 mg·g^-1respectively by HPLC.2.Predicted the internal mechanism of Prunella vulgaris L.stem and leaf total phenols to play an anti-inflammatory role,which may play an anti-inflammatory role through regulating PD-1,IL-17,AGE-RAGE and other signal pathways by acting on multiple key targets such as TP53,STAT3,JUN,HIF1A,CTNNB1,CASP3 and TNF,After analysis by means of molecular docking and molecular dynamics simulation,it was found that the Active ingredient of Prunella vulgaris L.,rosmarinic acid and salviafoliaside,had great anti-inflammatory potential.3.Animal experiments show that Prunella vulgaris L.stem and leaf total phenol have significant therapeutic effect on pelvic inflammatory model rats,which can repair the pathological state of rats’uterus and reduce the inflammatory cytokine IL-1β,IL-6 and IL-18.Transcriptome results show that its mechanism may be related to the intervention of staphylococcus aureus infection,estrogen,leukocyte migration across endothelium,VEGF,Gn RH,platelet activation and other signal pathways,as well as the expression of Pik3r2,Mapk12 and other genes in the regulatory pathway.4.The metabonomics results showed that after treatment with Prunella vulgaris L.total phenol extract,the inflammatory related metabolic pathways such as amino acid,ascorbic acid and Aldaric acid metabolic pathways in pelvic inflammatory model rats tended to return to normal levels;Relevant metabolites such as Phenylalanine,Threonine,serine and D-nenenebc glucuronic acid tend to return to normal levels,indicating that Prunella vulgaris L.can improve the abnormality of serum metabolites in inflammatory rats.Conclusion:In this thesis,the best extraction process of Prunella vulgaris L.stem and leaf total phenols was established through response surface model,and the chemical components of Prunella vulgaris L.stem and leaf total phenols were identified by UPLC-Q-TOF-MS/MS,and the content of key components was determined by HPLC.This part of research provides a reference for the basic research of Prunella vulgaris L.stem and leaf total phenols anti pelvic inflammatory substances;The anti-inflammatory mechanism of Prunella vulgaris L.stem and leaf total phenols was predicted by bioinformatics analysis;The anti-inflammatory effect of Prunella vulgaris L.stem and leaf total phenols was verified by pelvic inflammatory rat model,and key differential genes,differential metabolites and pathways were targeted through transcriptome and metabonomics,providing a reference for further in-depth research on the anti-inflammatory mechanism of Prunella vulgaris L..