To Explore The Mechanism Of Compound Jiegu Tablets Promoting Tendon-bone Healing After ACL Rupture And Reconstruction Based On Network Pharmacology

Objective: To analyze the effective chemical components of Compound Jiegu Tablets to study its mechanism of promoting tendon healing after anterior cruciate ligament rupture and reconstruction.Methods: Using Traditional Chinese Medicine Systems Pharmacology TCMSP,ETCM database,Pub Chem database and SIB database to search and screen the chemically active substances of Compound Jiegu Tablets and target proteins that act on the human body.At the same time,OMIM database and Gene Cards database predicts the target of NAFLD disease,and queries and corrects the protein name and corresponding gene name through the Uni Prot database,uses Cytoscape software to construct the active ingredient-target network diagram,builds the protein interaction network through the String database platform,and obtains it from the DAVID database GO(gene ontology)enrichment analysis and KEGG(Kyoto Encyclopedia of Genes and Genomes)enrichment analysis,use the Omic Share Tool cloud platform to visualize the results,and construct a target-pathway network diagram.Thirty-six Japanese white rabbits were selected as the tendon donor rabbits,and 9 rabbits were selected as the Dd group(sham operation group)according to the random number table method.The remaining 27 rabbits were all performed by the same operator before the operation Cruciate ligament severance,and modified anterior cruciate ligament reconstruction with processed allogeneic semitendinosus.After the rabbit is fully awakened from anesthesia,the rabbits are given a,b,and c three drugs by gavage,a,b The three drugs of,c are all configured by a third person who has nothing to do with this experiment to implement double-blind.They are divided into three groups A,B,and C at the same time.They are labeled as Aa,Bb,and Cc,and they are raised in separate cages.At 4,8,and 12 weeks after gavage,3 rabbits in each group were randomly selected and sacrificed in batches.Rabbit venous blood was collected for detection of PINP,TRACP-5b,C-RP,and complete blood white blood cell count;rabbit knee joint specimens were taken through Micro-CT observation of bone tunnel expansion rate,biomechanical detection of the ultimate tensile load of the ligament after transplantation,separate specimens for HE and Masson histological staining,and comparison of tendon-bone tunnel healing at each time point in each group.Result:After screening,one natural copper(calcined)active ingredient was finally included in this study,one active ingredient from soil beetle,one active ingredient from earthworm,32 active ingredients from mulberry bark,Drynaria fortunei(Preparation)15 active ingredients,8 Dipsacus active ingredients,2 Morus alba active ingredients,2 Angelica active ingredients,8 Boswellia active ingredients,45 Myrrh active ingredients,23 Angelica dahurica active ingredients,and safflower active ingredients There are 22 ingredients and 16 active ingredients in rhubarb.ACL breaks 177 disease targets,and obtains 41 drug-disease common target genes.Through the PPI protein interaction relationship,41 core genes were obtained,namely ITNF,VEGFA,MMP9,AKT1,PTGS2,CCL2,IL1 B,CASP3,TP53,MMP2,PPARG,EGF,MAPK1,CXCL8,SERPINE1,FOS,ESR1,SPP1,CCND1,MMP1,MAPK14,MMP3,HIF1 A,ICAM1,CAT,HMOX1,IFNG,CTNNB1,APP,ERBB2,RUNX2,IGFBP3,NOS2,COL1A1,GJA1,SOD1,IL1 A,LDLR,COL3A1,PTGS1,BCL2;GO enrichment analysis results Including cell response to lipopolysaccharide,inflammation,exosomes,membrane rafts,enzyme binding,protein binding,etc.;KEGG enrichment pathways include NF-κB signaling pathway,tumor necrosis factor signaling pathway,AMPK signaling pathway,etc.Confirmatory research results: The relative expression levels of PINP,TRACP-5b,C-RP,and total white blood cell count in the compound Jiegu Tablet group were(73.16±0.93ng/ml,9.84±2.02ng/ml,21.47±1.28ug/ml,4.36±0.38*10^9/L),osteopeptide tablet group(67.78±0.78ng/ml,10.79±0.04ng/ml,23.01±3.19ug/ml,4.49±0.09*10^9/L),placebo Group(69.82 ± 0.70ng/ml,10.47 ±0.27ng/ml,27.21 ± 5.32ug/ml,8.15 ±0.77*10^9/L),sham operation group(62.99±1.46ng/ml,10.59±1.02ng/ml,25.66±3.26ug/ml,4.42 ± 0.25*10^9/L),the relative expression of various indicators The amount of comparison between groups(F=17.88,p=0.000<0.01;F=0.73,p=0.546>0.05;F=2.080,p=0.134>0.05;F=16.883,p=0.000<0.01).Conclusion: The compound Jiegu tablet may regulate the osteoclast differentiation signal pathway by regulating ITNF,VEGFA,MMP9,AKT1 and other targets to promote tendon bone healing after ACL rupture and reconstruction.