Study On Mechanism Of Xiaoqinglong Decoction Syndrome Based On PI3K/Akt/mTOR Signal Pathway

Objection: To investigate the mechanism of TCM Syndromes of Xiaoqinglong(XQLD)Decoction basd on the PI3K/Akt/mTOR signal pathway,we predicted and screened the targets of XQLD and the possible mechanism of action of XQLD through network pharmacology and experimentally verify whether XQLD exerts its therapeutic effect through regulating PI3K/AKT signaling pathway.Methods: 1.Predict and screen the targets and the protential mechanisms of action of XQLD through network pharmacology: collecting the active compounds and thier targets of XQLD with the help of Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)platform;with the help of the Online Mendelian Inheritance in Man(OMIM)database and the genome annotation database platform(Gene Cards),we collected bronchial asthma 、chronic bronchitis and upper respiratory tract infection’s target,and we got the intersecting genes by the Venn diagram 2.1;the intersection genes were imported into String 11.0platform to construct a protein-protein interaction(PPI)network;use the DAVID 6.7database and Omicshare Tool software to perform Gene Ontology(GO)Analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG).2.Established a rat model of XQLD Syndrome,with a blank group,a model group,and high,medium,and low dose groups of XQLD,and observe the general state,body weight,and histomorphological changes of rat lung tissue.Measured the levels of tumor necrosis factor-alpha(TNF-α),interleukin-6(IL-6),and interleukin-8(IL-8)in rat serum by Enzyme-Linked Immuno Sorbent Assay(ELISA).Quantitative Realtime PCR was used to measure the relative levels of PI3 K,Akt,and mTOR m RNA in rat lung tissue.Western Blot was used to detect the protein levels of PI3 K,Akt,and mTOR in rat lung tissue.Results: 1.We obtained 169 effective compounds,1105 drug targets,and 824 disease targets of XQLD and 254 potential targets.By constructing a "compound-target"network diagram,we got 110 core components and 106 core targets of XQLD’s treatment.The core ingredients include kaempferol,beta-sitosterol,quercetin,naringenin,Stigmasterol,Jaranol,and sitosterol.sitosterol),baicalein,Glypallichalcone,Deoxyharringtonine.The core targets include ESR1、ESR2、ACHE、CDK2、EGFR、ABCG2ABCB1、CCNB1、MMP13、MMP3,etc.KEGG enrichment analysis showed that the potential pathways were PI3K-Akt signaling pathway,mTOR signaling pathway,HIF-1 signaling pathway,Erb B signaling pathway,Ras signaling pathway,Fox O signaling pathway,Rap1 signaling pathway,VEGF signaling pathway,and TNF signaling pathway.2.General results of rats: During the modeling process,one rat died in the model group,and the rest of the groups were successfully modeled.relatived with rats in the blank group,rats in other groups showed cough,shortness of breath,wheezing,sneezing,reduced diet,slow response,slow movement,love to get together,standing hair,arched back,curled up,etc.Changes in body weight of rats: Compared with the blank group,the body weight of rats in the model group and the XQLD treatment group did not increase significantly after modeling,and even showed negative growth(P<0.01);after drug intervention treatment,the body weight of rats still did not increase significantly compared with the blank group(P>0.05).Compared with the model group,the body weight of rats in XQLD treatment groups increased significantly(P<0.01).The lung histopathological manifestations of rats: there was no obvious abnormality in the lung tissue structure of rats in the blank group,the alveolar structure was regular and intact,and the alveolar wall structure was clear,and there was no obvious inflammatory cell infiltration.In the model group,the alveolar septum was widened,the alveolar cavity was narrowed or even collapsed,some of the alveolar cavities were dilated,and the alveolar interstitium and bronchial wall were infiltrated by inflammatory cells,mostly lymphocytes and eosinophils.After the administration of XQLD,the collapse of alveolar lumen narrowing,widening of alveolar interstitium and reduction of inflammatory cell infiltration were improved in the XQLD high-dose group medium-dose group,but it was difficult to restore the structural state of the lung tissue in the blank group.RT-q PCR results showed that compared with the blank group,the relative expression of PI3 K,AKT,and mTOR m RNA in the model group increased(P<0.05);compared with the model group,the relative expression of PI3 K,Akt,and mTOR m RNA in the XQLD high-dose group Decrease(P<0.05 or P<0.01);Compared with the model group,the expression of PI3 K,Akt and mTOR m RNA in the XQLD low-dose group decreased,but there was no statistical significance(P>0.05).ELISA results showed that compared with the blank group,the serum levels of TNF-α,IL-6 and IL-8 in the model group increased(P<0.05);compared with the model group,the XQLD medium-dose group and XQLD were higher The content of TNF-α,IL-6,and IL-8 in the serum of rats in the dose group decreased(P<0.05);compared with the model group,the content of TNF-α,IL-6 and IL-8 in the serum of rats in the XQLD low-dose group decreased,But not statistically significant(P>0.05).Western blot results showed that compared with the blank group,the protein content of PI3 K,AKT,and mTOR in the lung tissue of the model group increased(P<0.05);compared with the model group,the medium dose of XQLD group and the high dose of XQLD The protein content of PI3 K,AKT,and mTOR in the lung tissue of rats in the group decreased(P<0.05);compared with the model group,the content of PI3 K,AKT,and mTOR protein in the lung tissue of rats in the XQLD low-dose group decreased,but no statistics Significance(P>0.05).Conclusion:(1)Based on the network pharmacology,this study predicted the target of XQLD Syndrome and obtained 824 disease targets of XQLD Syndrome,110 core components,and core targets of XQLD.106.The mechanism of action of XQLD is related to PI3K-Akt signaling pathway,mTOR signaling pathway,HIF-1 signaling pathway,Erb B signaling pathway,Ras signaling pathway,Fox O signaling pathway,Rap1 signaling pathway,VEGF signaling pathway,and TNF signaling pathway.(2)The development of XQLD Syndrome is associated with inflammatory and immune responses regulated by PI3K/Akt/mTOR signaling pathway,and XQLD can improve lung inflammation by downregulating the expression of protein and m RNA of PI3 K,AKT,and mTOR,inhibiting pro-inflammatory factors in serum and infiltration of inflammatory cells in lung tissue,most significantly in the high-dose group.