Study On The Mechanism Of Total Flavonoids Of Litchi Seed Against Liver Fibrosis In Rats Based On Proteomics

Objective: To observe the mechanism of total flavone of Litchi chinensis Sonn on HSC-T6 cells against liver fibrosis in vitro,and provide reference for its further development and utilization.Methods: After HSC-T6 was interfered with by litchi nuclear total flavonoids and litchi nuclear total flavonoids serum for 14 days,protein was extracted,differential protein was screened by protein quantitative DIA and DDA technology,and differential protein was enriched and annotated by GO database,signal transduction pathway analysis and protein interaction analysis by KEGG database.The TCMSP database was used to screen the effective components of TFL,and the protein-protein interaction network of TFL chemical components was constructed in STRING10.5 database.The related targets of TFL and liver fibrosis were obtained by Gene Cards database and analyzed by bioinformatics.Results: 6011 differential proteins,168 up-regulated proteins and 203 down regulated proteins were identified in total flavonoids group and control group.Through GO analysis,differential proteins mainly participate in binding,catalytic activity,molecular function regulators,transcriptional regulatory activity and other molecular functions.Cell components mainly exist in the cell,organelle,extracellular,cell membrane and mainly participate in 27 biological processes,such as cell process,metabolic process,biological regulation,regulatory biological process.Through Pathway analysis,we found that it mainly involved 30 signaling pathways,such as metabolicpathway,complement and coagulation cascade,transcription disorder in tumor,NF-? B signaling pathway,PPAR signaling pathway,HIF-1 signaling pathway.It was found that the up-regulated protein was mainly distributed in extracellular,mitochondrial,peroxidase,endoplasmic reticulum and cytoplasmic sol-peroxidase,and the down regulated protein was mainly distributed in nucleus,cytoplasmic sol,cytoplasmic membrane and cytoplasmic sol-nucleus.Two active components were screened from TFL:epicatechin and quercetin,and 72 potential targets.After checking and comparing with the targets of liver fibrosis related pathogenesis,15 target genes of TFL and liver fibrosis related pathogenesis were obtained,including MMP2(matrix metalloproteinase)and CCL2(chemokine CC)Subfamily),TP53(tumor suppressor protein),HMOX1(heme oxygenase 1),IFNG(interferon ? gene),etc.The above targets are mainly distributed in the intercellular matrix,secretory granules and protein extracellular matrix,and play a therapeutic role in liver fibrosis mainly through the molecular functions of immunoreaction,positive regulation of chemokine biosynthesis process,positive regulation of T cell proliferation,positive regulation of RNA polymerase II promoter transcription,etc.There are 32 signal pathways involved in the above targets,among which 9 are related to liver fibrosis,including tumor signal pathway,HIF-1 signal pathway,interaction between cytokines and cytokine receptor,TNF signal pathway,micro RNA in cancer,PI3 K Akt signal pathway and nod(nucleotide binding oligomerization domain)Like receptor signaling pathway,NF-? B signaling pathway and MAPK signaling pathway.IL1B(recombinant human interleukin-1 beta),IL6(interleukin-6),IFNG and other genes in the "component target pathway" network of TFL for the treatment of liver fibrosis are the core targets in the TFL effective component treatment of liver fibrosis network(nodal value ?15,median centrality > 0.8).Tumor signaling pathway,TNF signalingpathway,interaction signaling pathway between cytokines and cytokine receptor are the core action pathways of TFL active ingredients in the treatment of liver fibrosis(nodal value ? 5,median centrality > 0.001).Conclusion: Protein quantitative DIA and DDA can quickly screen out differential proteins,and GO analysis and KEGG analysis can explain the mechanism of TFL's anti fibrosis in vitro in a holistic view;HMOX1,NQO1,ACACA and CDK1 can play an anti fibrosis role when they are abnormally expressed.It can be inferred that the mechanism of anti fibrosis of TFL in litchi seed may be related to the expression of these four differential proteins,which may be the target proteins of TFL in litchi seed.