A Preliminary Study On The Metabonomics And Molecular Mechanism Of Qingyi Decoction In The Treatment Of Acute Pancreatitis In Mice

Objective:1.To establish the quality standard of Qingyi Decoction internal control by HPLC;2.Based on metabolomics and RT-PCR technology,the molecular mechanism of Qingyi Decoction on acute pancreatitis was explored.Methods:?1?The 10 active ingredients of 5 herbs in Qingyi Decoction were selected and analyzed by HPLC.The accuracy,specificity,linearity,precision,repeatability and stability of the method were verified.?2?To establish the model of acute pancreatitis in mice with different dose of model making agent.To study the effect of Qingyi Decoction on acute pancreatitis in mice,the pathological changes of pancreas,lung and small intestine,as well as biochemical and inflammatory indexes such as AMS,Lip,SOD and IL-6 were taken as the main research object.?3?The UPLC-Q-TOF MS technical analysis between groups serum samples,the data analysis of PCA and OPLS-DA,then use HMDB?http://www.hmdb.ca/?retrieved from preliminary screening of potential biomarkers,reuse MetaboAnalyst?https://www.metaboanalyst.ca/?andKEGGpathwayanalysis?https://www.genome.jp/kegg/?.?4?Detection of TNF-?,COX-2,NOX2,MMP-9gene expression in pancreatic tissue by RT-PCR.Result:?1?The paeoniflorin,picroside II,picroside I,baicalin,baicalein,aloe-emodin,rhein,saikoside B2,emodin and chrysophanol 10 active ingredients in Qingyi Decoction can be completely separated,and the linear relationship was good within a certain concentration range.The precision,stability and repeatability were in line with the analysis of the quality standard of the drug in the 2015 edition of the Pharmacopoeia of the People's Republic of China?RSD?2.0%?.The content of 10active ingredients in the three batches of Qingyi Decoction were between251.22²76.12,649.52⁶94.33,242.74³06.69,935.55⁹79.03,33.86³5.97,8.31⁹.65,15.17²4.23,6.74⁹.74,4.16⁷.47,8.39⁹.27 mg respectively.?2?It was finally determined that 80?g·kg^-1 cerulein and 12 mg·kg^-1lipopolysaccharide were used for modeling.The levels of serum AMS,Lip and IL-6in the model group were significantly higher than those in the blank group,and the level of SOD was significantly decreased?P<0.01?.Compared with the model group,the pathological sections of pancreas,lung and small intestine of Qingyi Decoction group improved obviously and became normal;serum AMS,Lip and IL-6 levels were significantly decreased,and SOD level was significantly increased.It shows that Qingyi Decoction has a significant effect on acute pancreatitis.?3?Compared with normal mice,there were significant changes in 28 potential metabolites in mice with acute pancreatitis,mainly involving Linoleic acid metabolism,Taurine and hypotaurine metabolism and Arachidonic acid metabolism.After giving Qingyi Decoction,there were 17 recalls with different degrees of callback,indicating that Qingyi Decoction can relieve acute pancreatitis in mice by interfering with linoleic acid metabolism,taurine and hypotauric acid metabolism and arachidonic acid metabolic pathway.?4?Compared with the blank group,the relative expression levels of TNF-?,COX-2,NOX2,MMP-9 mRNA in the model group were significantly increased.Compared with the model group,the relative expression levels of TNF-?,COX-2,NOX2,MMP-9 mRNA in pancreatic tissue were significantly decreased.Conclusion:We successfully established the content determination method of10 index components in Qingyi Decoction,providing experimental data guarantee for ensuring the effectiveness of drugs.Qingyi Decoction can alleviate the acute pancreatitis induced by hylocinin combined with lipopolysaccharide in mice.Serum metabolomics studies showed that it mainly plays a role in regulating the metabolic pathways of linoleic acid,taurine,taurine and arachidonic acid.RT-PCR showed that the drug could inhibit the expression of TNF-??COX-2?NOX2?MMP-9,which are related to inflammation in the pancreatic tissue of mice.