Based On Network Pharmacology And Combined With Experiments To Explore The Anti-inflammatory Mechanism Of Tianshen Capsule

Objective: Inflammation is an important part of the body’s immune function.Mild or short-term inflammation is an important defense against external stimuli,while severe or long-term inflammation can cause damage to the body and even lead to diseases.There are a large number of oral and maxillofacial diseases related to inflammation,such as pericoronitis of wisdom teeth,osteomyelitis of jaw,periodontitis and some oral mucosal diseases such as lichen planus.In the treatment of these diseases,controlling inflammation is an important means.Traditional Chinese medicine plays an important role in the treating inflammation and inflammation-related diseases,and has a broad research prospect.Tianshen capsule is a new drug for the treatment of cardiovascular diseases,which contains a variety of traditional Chinese medicine ingredients and has a certain anti-inflammatory effect,but because of its multiple components,complex chemical components and corresponding disease targets,it is difficult to directly use experiments to explore the related mechanism of its anti-inflammatory effect.Based on systems biology,network pharmacology is a new technology that uses retrieval and database analysis to build a network and analyze,which can be effectively applied to the research and development of traditional Chinese medicine.The anti-inflammatory mechanism of Tianshen capsule was analyzed by the method of network pharmacology and further verified by in vitro experiments.Methods: The specific chemical components of traditional Chinese medicine components of Tianshen capsule were found by TCMSP and other databases,and the effective components were further screened by Oral Bioavailability（OB）value and Drug-Likeness（DL）value.TCMSP and Swiss Target Prediction databases were used to query or predict the corresponding targets of effective components,and Gene Cards and Dis Ge NET databases were used to query inflammation-related targets,and get the common targets of component targets and disease targets.The PPI network analysis of the common target is carried out by using String database,and the core target is selected.DAVID database,Bioconductor data package and R software were used for Gene Ontology（GO）and（Kyoto Encyclopedia of Genes and Genomes）KEGG analysis of the core targets,and Discovery Studio software was used for further molecular docking.Raw264.7 cells were induced by Lipopolysaccharide（LPS）to establish inflammatory model,and different concentrations of water extract of Tianshen capsule were used to intervene.Griess method was used to detect the changes of NO in the supernatant,ELISA method was used to detect the expression of TNF-α and IL-6 in the supernatant,and the relative expression of p-JAK2 and p-STAT3 was detected by Western blot.The results of network pharmacology analysis were further verified by experiments.Results: A total of 855 chemical constituents of Tianshen capsule were found by using multiple databases,and 147 effective components were obtained by screening with OB ≥ 30% and DL ≥ 0.18 as the lowest conditions.A total of 560 targets corresponding to effective components were obtained by TCMSP and Swiss Target Prediction.The topological analysis of PPI network shows that Tianshen capsule can mainly act on STAT3,TP53,JUN,AKT1,SRC,MAKP1 and other targets.GO analysis showed that it mainly acted on DNA-binding transcription binding,RNA polymerase II-specific DNA-binding transcription binding and ubiquitinlike protein ligase binding.KEGG pathway analysis showed that it may play an anti-inflammatory effect through PI3K-AKT signal pathway,MAPK signal pathway,Toll receptor signal pathway,JAK/STAT signal pathway and so on.The in vitro cell experiment showed that the water extract of Tianshen capsule could significantly reduce the expression of NO,TNF-α and IL-6（P<0.05）.The results of Western blot showed that the water extract of Tianshen capsule could significantly inhibit the expression of p-JAK2 and p-STAT3 protein and inhibit JAK2/STAT3 signal pathway（P<0.05）.Conclusion: The network pharmacological analysis showed that Tianshen capsule plays an anti-inflammatory role mainly through STAT3,TP53,JUN and other targets,JAK/STAT,PI3K-AKT and other signal pathways.Cell experiments also proved that the water extract of Tianshen capsule could inhibit the inflammation induced by LPS in Raw264.7 cells by inhibiting JAK2/STAT3 signal pathway.