Study On The Formation Mechanism Of Primary Choledocholithiasis And The Treatment Mechanism Of Cholelithiasis With Bupleurum

Experiment 1: Comparison of anatomical structure of common bile duct between primary and secondary cholelithiasis Objective: To investigate the effect of common bile duct dilatation and angle formation on choledocholithiasis.Methods: A case-control study was used in this study.From January 2019 to December 2019,patients with choledocholithiasis treated by minimally invasive surgery in Nankai Hospital of Tianjin were selected in this study.According to the inclusion and exclusion criteria,they were divided into primary choledoch olithiasis group(37 cases)and secondary choledocholithiasis group(38 cases).Prospectively collect the clinical data of patients(gender,age,basic symptoms of admission,history,blood routine,liver function,diameter of common bile duct,angle of common bile duct,maximum stone diameter,number of stones,etc.)and compare the difference between the two groups.Results: there was no difference in the angle of common bile duct between the primary choledocholithiasis group and the secondary choledocholithiasis group (128.00(113.00-141.00)vs 125.00(117.00-135.00),z =-0.212,P = 0.832);the diameter of common bile duct of the primary choledocholithiasis group(1.11(0.82-1.41)cm)was larger than that of the secondary choledocholithiasis group(1.00(0.73-1.39)cm),the difference was statistically significant(t = 3.486,P = 0.001 < 0.05).Conclusion: the primary choledocholithiasis is related to cholestasis caused by choledochal dilatation.Experiment 2: Non target metabonomics analysis of the differences of bile components between primary and secondary choledocholithiasis Objective: To explore the difference of bile components between primary and secondary choledocholithiasis by non-target metabonomics,and to explore the formation mechanism of choledocholithiasis,so as to provide a new clinical idea for the treatment of choledocholithiasis.Methods:In the early morning of the second day after ERCP,bile was collected from the patients through nasobiliary duct before breakfast.Five bile samples were selected from each group by random sampling.The bile was separated and identified by ultra-high performance liquid chromatography(UPLC)and Q-TOF mass spectrometry,and the differential metabolites were screened by PCA,PLS-DA,OPLS-DA.Results:Comparison of bile metabolites: the response intensity and retention time of the peaks of total ion flow mass spectrometry(TIC)in the positive and negative ion detection mode of QC samples overlap approximately.The variation caused by instrument error is small and the data quality is reliable.In the PCA model,QC samples are closely gathered together,and the repeatability of this experiment is good.OPLS-DA model can distinguish two groups of samples.Twenty metabolites with significant difference were detected,including dihy droxyacetone,caprylic acid,lauric acid,2-Naphthalenesulfonic acid,α-n-phen ylacetyl-l-glutamine,pregnenolone sulfate,17-β-estradiol glucosamine,urea,carnitine,phenylalanine isoleucine,25 hydroxyvitamin D3,etc.Conclusion: There may be a potential metabolic mechanism that affects the for mation and recurrence of primary choledocholithiasis.Experiment 3: Explore the mechanism of Bupleurum in the treatment of cholelithiasis based on network pharmacology Objective: To explore the effective components and targets of Bupleurum in the treatment of gallstones,and to analyze the mechanism of Bupleurum in the treatment of gallstones,so as to provide a scientific basis for Bupleurum in the treatment of gallstones.Methods:We used TCMSP to find the effective components and related targets of Bupleurum;By typing "Cholelithiasis" into Gene Cards,we found genes rela ted to cholelithiasis;The regulatory network of Bupleurum chinense and cholelithiasis was obtained by using Cytoscape 3.7 software,"string" was used to analyze the interaction between drug and common target gene related protein,and PPI network was obtained.Bioconductor was used to change the gene name of drug and gene target intersection into gene ID;GO enrichment analysis and KEGG enrichment analysis are carried out by running relevant procedures in r3.6.1。 Results: There are 53 same target genes in Bupleurum and cholelithiasis;There are 10 kinds of components in Bupleurum that have the effect of treating gallstones,which are linolyl acetate,stigmasterol,isorhamnetin,kaempferol,3,5,6,7-tetramethoxy-2-(3,4,5-trimethoxyphenyl)chromone,areapillin,cubebin,longikaurin a,petunidin and quercetin;the most adjacent nodes in PPI network diagram are IL6;in GO(gene ontology)enrichment analysis,receiver life activity enrichment is the most significant;in KEGG(Kyoto Encyclopedia of genes and genes)enrichment analysis,five path diagrams are obtained,namely AGE-R AGE、Bladder cancer、Fluid shear stress and atherosclerosis、HIF-1、Endocrine resistance,AGE-RAGE is the most significant.Conclusion: the mechanism of Bupleurum in the treatment of cholelithiasis is complex,which may be treated by anti-inflammatory,anti-oxidation,cholesterol regulation and carbohydrate metabolism.