| StudylObjective:to systematically evaluate the effective active chemical components,pharmac odynamic genes and main pathways of Yifei Sanjie jiedu decoction in the treatment of lung cancer.Methods:the chemical constituents and drug action targets in Yifei Sanjie jiedu decoc-tion were searched through the pharmacological database of traditional Chinese medici ne system,and the active chemical components and drug effective targets in Yifei San jie jiedu decoction were screened according to the conditions of "bioavailability OB≥30%and drug-like DL≥ 0.18".The target gene prediction website human gene databa-se(Genecards)and human Mendelian genetic database(OMIM)were used to predict and screen the targets for the treatment of lung cancer.With the help of RMel 3.6.2 development tool to find the intersection of Yifei Sanjie jiedu decoction action target and lung cancer related target,get the efficacy gene of Yifei Sanjie jiedu decoction in tervention on lung cancer,draw Venn diagram,directly reflect the efficacy gene.By u-sing R-3.6.2 tool and bioconductor database,the regulatory network construction of tr-aditional Chinese medicine,the enrichment analysis of GO gene function and KEGG pathway and network visualization were carried out to reflect the potential relationship among drug,gene and disease,and the main signal pathway of Yifei Sanjie jiedu de-coction on lung cancer was obtained.Results:1.The results of network pharmacology research showed that through the trad itional Chinese medicine system pharmacology database and analysis database,accordin g to the condition of "OB≥ 30%and DL≥ 0.18",a total of 95 active chemical constit uents were obtained,including quercetin,digitalis flavone,sitosterol(beta-sitosterol),k-aempferol,stigmasterol,isorhamnetin,baicalin,baicalin,anthraquinones,8-isopentenyl-k-aempferol,The main active chemical constituents of 7-hydroxy-5pyrine-8-dimethoxy-2-phenyl-chromone,Heindrin,farnesin,polysaccharides and coumarin were active.1083 e-ffective targets were obtained,22184 lung cancer-related gene targets were obtained b-y Gencard and OMIM database,and 215 drug-related genes were obtained,including TP53,BCL-2,BIRC5 and other genes.Through the functional analysis of the GO gen-e,the functional enrichment of the gene includes selective and non-covalent interactio-n with a specific deoxyribonucleic acid sequence in the regulatory region of the ribon-ucleic acid polymerase 2 transcriptional gene to activate or increase transcription(DN A-binding transcription activator activity,RNA polymerase Ⅱ-specific,25/215)and intera-ct with the receptor to affect the active(receptor ligand activity,20/215 of the gene p-roduct with changed receptor activity.And cytokine receptor binding to(cytokine rece-ptor binding,19/215),which directly affects the cytokine activity(cytokine activity,17/215).Through the KEGG pathway enrichment analysis,according to the first 20 pathw-ays with the highest enrichment degree in the P value map,in-depth analysis showed that the pharmacological genes of Yifei Sanjie jiedu decoction were mainly enriched i-n the PI3K-AKT,AGE-RAGE,MAPK signal pathway.Conclusion:after the analysis of the research methods of network pharmacology,we f-ound that Yifei Sanjie jiedu decoction contains a variety of effective chemical compon-ents to inhibit lung cancer,and generally speaking,flavonoids have the most targets.To a certain extent,it can reflect that flavonoids play an important role in the process of anti-lung cancer.One hundred and twenty-five effective genes of Yifei Sanjie jied u decoction in the treatment of lung cancer were screened,including TTP53,BCL-2,BI RC5 and other hot genes.The functional analysis of GO gene showed that Yifei Sanji e jiedu decoction could treat lung cancer by regulating the activity of cytokines and r-eceptor ligands,affecting the binding of cytokine receptors and interacting with specifi-c DNA in the regulatory region of ribonucleic acid polymerase 2 transcriptional gene.KEGG pathway enrichment analysis shows that Yifei Sanjie jiedu decoction plays a r-ole in lung cancer through multi-channel,multi-aspect and multi-target,mainly throug-h PI3K-AKT,AGE-RAGE,MAPK signal pathway to induce or promote cell apoptosis and inhibit proliferation,so as to achieve anti-tumor effect.Study 2Objective:to explore the effect and mechanism of Yifei Sanjie jiedu decoction on human lung adenocarcinoma A549 cells in combination with the results of network pharmacology.Methods:SD rats were perfused with Yifei Sanjie jiedu decoction,and the control group was given the same volume of normal saline.After intragastric administration for 5 days,abdominal aorta blood was taken and placed in anticoagulant tube to obtain drug-containing plasma and blank plasma respectively.Human lung adenocarcinoma A549 cells in the stage of growth and division were selected and treated with different concentrations of blank plasma.after 24 hours of treatment,the cell viability was detected by CCK8 method,and the cell inhibition concentration of blank plasma was obtained,thus the range and gradient of drug-containing plasma concentration gradient were determined.Different concentrations of drug-containing plasma of Yifei Sanjie jiedu decoction were added to A549 cells,the same concentration of blank plasma was added in the control group,and no plasma was added in the routine control group.After 24 hours and 48 hours of treatment,the morphological changes of cells were observed under inverted microscope,the inhibition of cell proliferation was detected by CCK8 method,and the expression of TP53,BCL-2 and BIRC5 mRNA in human lung adenocarcinoma A549 was detected by RT-PCR method.Compared with the control group,the experimental data were compared with the control group.The experimental data were compared and analyzed by independent sample t-test,and the obtained data were expressed by SD(independent X±S).Results:the results of blank plasma proliferation inhibition experiment showed that there was no significant difference in the inhibitory effect between the blank plasma group and the routine control group(P<0.05)in the range of 0%murine 10%concentration(P<0.05),which was a safe range.In order to eliminate the inhibitory effect of blank plasma on cells and affect the experimental results,the concentration gradient of drug-containing plasma was set to 2.5%,5%,7.5%,7.5%,10%,and the morphology of cells at 24 hours and 48 hours was observed under inverted microscope.blank plasma group:lung adenocarcinoma cells were epithelial,adherent growth,cell shape was oval and regular,uniform,tight connection between cells;In the drug-containing plasma group:with the extension of the culture time and the increase of the concentration of the drug-containing plasma,the cell morphology changed obviously,the outline enhanced,the volume shrank,the shape was irregular,or flattened or extended,the intercellular space became larger,and some cells became round and floated.CCK8 detection showed that different concentrations of drug-containing plasma inhibited the proliferation of lung adenocarcinoma A549 cells for 24 h and 48 h.After statistical analysis,compared with the corresponding concentration of blank plasma group at the same time,the inhibitory effect was stronger with the increase of drug-containing plasma concentration,and the inhibitory effect was gradually enhanced with the extension of action time(P<0.05).RT-PCR results showed that the expression of BCL-2 and BIRC5 mRNA in lung adenocarcinoma A549 cells decreased with the change of drug-containing plasma concentration,while the expression level of TP53 mRNA increased gradually.After statistical analysis,compared with the blank plasma group,there were significant differences in BCL-2,TP5 3 and BIRC5 gene expression in different concentrations of medicated plasma group(P<0.05).Conclusion:Yifei Sanjie jiedu decoction can inhibit the proliferation of human lung adenocarcinoma A549 cells,and its mechanism may be related to inhibiting the expression of BCL-2 and BIRC5 mRNA and promoting the expression of TP53 mRNA. |
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