Proteomics Annotate Therapeutic Effect Of Xiaoai Jiedu Decoction In Hepatocellular Carcinoma Rats

Objective:TCM Carcinogenic Pathogenesis theory "cancer toxins" propose the view that "cancer toxins" plays a vital role in the development of carcinoma.In this theory,qi stagnation and phlegm-blood stasis,heat-toxin mainly contribute to the formation of cancer toxins,Xiaoai Jiedu decoction is a prescription which based on the theory of cancer toxins.This experiment mainly use proteomics to detect the differentially expressed proteins in liver tissue and serum sample of Hepatocellular carcinoma(HCC)rats in Xiaoaijiedu decoction group,model group and normal group.We aim to explore the potential mechanism of Xiaoaijiedu decoction on Hepatocellular carcinoma.Methods:40 male SD rats were randomly divided into normal group,model group,Xiaoai Jiedu low-does(LXAJD)group,Xiaoai Jiedu high-does(HXAJD)group,10 in each group.Rats in the model group?LXAJD group?HXAJD group were treatd with 0.01%N-Nitrosodiethylamine(DEN)solution drinking to establish a hepatoma model.At the 4th week,rats in the LXAJD?HXAJD group were given a dose of 10.87 or 21.74g.kg-1.d-1 of XAJD water extract by gavage respectivey,normal and model groups were given the same volume of saline.Execute all rats at the end of the 16th week,take the liver samples and serum samples;Prepare the protein samples.All samples were labeled with TMT peptides and identified by mass spectrometry.GO analysis and pathway analysis of differentially expressed proteins was performed by KOBAS database,String database was used for PPI analysis.Results:(1)Differentially expressed proteins in liver tissueA total of 6287 protein spots were identified.Compared with the normal group,1002 proteins were differentially expressed in the model group,466 proteins were up-regulated and 536 proteins were down-regulated.Compared with the model group,456 proteins were differentially expressed in the LXAJD group,including 265 up-regulated proteins and 191 down-regulated proteins.752 proteins were differentially expressed in the HXAJD group,including 424 up-regulated proteins and 328 down-regulated proteins.All the differentially expressed proteins are mainly involved in metabolism,glycolysis,immune responses,inflammatory responses,heat shock,oxidative stress,reactive oxygen species,cell cycle,apoptosis,cell proliferation,cell differentiation,cell migration,cell morphology,blood vessel growth,Matrix Metalloproteinases,Cell Adhesion Molecules,Fibrin Extracellular Matrix Proteins,Gap Junction Proteins,Plasminogen Receptors,Epithelial Barrier Function,Transcription Factors,Liver Regeneration and Repair,Oncogene,etc.Pathway analysis revealed that all differentially expressed proteins involved in the following pathways(according to the enrichment scores):PPAR pathway,glycolysis,and AMPK pathways;Extracellular matrix receptor interaction pathway,plasminogen activator pathway,DNA replication,cell cycle,cell adhesion molecule,PI3K-Akt signaling pathway,p53 signaling pathway,Jak/STAT signaling pathway,gap junctions,TGF-? signaling pathway,autophagy regulation,Wnt signaling pathway,HIF-1 signaling pathway,apoptotic signaling pathway,cadherin signaling pathway,B cell activation pathway,FoxO signaling pathway,Nuclear transcription factor-?B(NF-?B)signaling pathway,etc(2)Differentially expressed proteins in serumA total of 1151 protein spots were identified.Compared with the normal group,234 proteins were differentially expressed in the model group,including 151 up-regulated proteins and 83 down-regulated proteins.Compared with the model group,144 proteins were differentially expressed in the XAJD low-dose group,including 56 up-regulated proteins and 88 down-regulated proteins.182 proteins were differentially expressed in the XAJD high-dose group,including 94 up-regulated proteins and 88 down-regulated proteins.These differentially expressed proteins are mainly involved in metabolism,inflammation,immune regulation,heat shock,oxidative stress,cell cycle and apoptosis,cell growth and differentiation,angiogenesis,cancer-related,liver regeneration repair.Pathway analysis revealed that all differentially expressed proteins involved in the following pathways(according to the enrichment scores):PPAR signaling pathway ECM-receptor-interaction signaling pathway,TLRs signaling pathway?HIF-1 signaling pathway?p53 signaling pathway?PI3K-Akt signaling pathway?mTOR signaling pathway?Phagosome?NF-?B signaling pathway?Glycolysis,TGF-? signaling pathway,B cell activation,Jak/STAT signaling pathway,FOXO signaling pathway,etc.Conclusion:We applied proteomics to detect the differentially expressed proteins in the rats of Normal,model or XAJD group to explore the potential mechanism of XAJD.We found that the main anti-HCC mechanism of XAJD include regulation of metabolic and ECM-receptor interaction,the regulation of the inflammatory response in tumor microenvironment also contribute to it.It shows that XAJD has multi-target and multi-pathway regulating effect.