Immune Infiltration Analysis and Target Prediction of Traditional Chinese Medicine in Nonalcoholic Fatty Liver and Liver Fibrosis
Objective This study explores the correlation between macrophage markers and NAFLD and liver fibrosis,in order to further explore the pathogenesis of NAFLD liver fibrosis,explore effective components of traditional Chinese medicine,and provide help and reference for the prevention and treatment of liver fibrosis by traditional Chinese medicine.Methods Utilizing bioinformatics methods,NAFLD gene chips were analyzed to identify differentially expressed genes.Through WGCNA co-expression network analysis and LASSO model construction,core expression genes were selected.CIBERSORT was employed for immune cell infiltration analysis,exploring the correlation between macrophage phenotypes and differentially expressed genes,as well as the degree of fibrosis.External datasets were used to identify relevant biomarkers for macrophages in NAFLD and liver fibrosis.Reverse transcription-polymerase chain reaction(RT-PCR)was conducted to validate the expression differences of core genes across different macrophage subtypes.Additionally,the Coremine medical database was utilized for ontology-based information retrieval to screen and analyze traditional Chinese medicine treatment targets for NAFLD.Results A total of 15 core genes related to NAFLD and liver fibrosis were screened out in this study.CIBERSORT algorithm analysis results showed that the occurrence of NAFLD was related to various immune cells,among which M1 macrophages were significantly elevated in the NAFLD group,with a significant difference(P<0.01).M1 macrophages demonstrated a positive correlation with the degree of liver fibrosis,while M2 macrophages exhibited a negative correlation.The correlation analysis between core genes and macrophages revealed that KDM4A,MED30,IKBKG,ENO1,and HSPA8 were negatively correlated with total macrophage count.The expression of IL-6,FOS,EDN2,and NR4A2 displayed a negative correlation with M1 macrophage count,whereas it showed a positive correlation with M2 macrophage count.The results of external dataset validation and in vitro RT-PCR validation indicated that the expression levels of FOS and NR4A2 mRNA in M2 macrophages were higher than those in M1 macrophages(P<0.001).This expression was negatively correlated with M1 macrophages and positively correlated with M2 macrophages.Based on the core target through Coremine medical database,according to the drug efficacy and meridian,we screened out the related traditional Chinese medicine such as Salvia miltiorrhiza,Cortex moutan,Radix Rehmanniae,Angelica sinensis,Alisma,Semen Hoveniae,Ginseng,Dogwood,Hawthorn and so on.The efficacy analysis of these Chinese medicines mainly focused on clearing heat and detoxification,removing dampness and removing phlegm,activating blood circulation and removing blood stasis.Among these,Cortex moutan was identified as the most frequently selected medicine.Conclusion Liver macrophages are target cells for treating liver diseases.Regulating the M2 polarization of macrophages contributes to the treatment of NAFLD and related liver fibrosis.c-Fos and NR4A2 are potential key target points,and traditional Chinese medicines like Moutan Cortex have hepatoprotective effects.This provides new insights for further researching the pathogenesis and treatment methods of NAFLD and liver fibrosis.
Nonalcoholic fatty liverLiver fibrosisMacrophages markersImmune infiltrationTarget in Chinese herbs
NETL
NSTL
万方数据
