To Explore the Mechanism of Xiaozhongzhitong Mixture in the Treatment of Ischemia-Reperfusion Injury of Skin Flap Based on TMT Proteomics / 世界科学技术-中医药现代化

ABSTRACT

Objective Tandem Mass Tag(TMT)in vitro quantitative proteomics technology was used to study the mechanism of Xiaozhongzhitong Mixture in the treatment of rat skin flap ischemia-reperfusion injury(FIRI).Methods Thirty SD rats were randomly divided into a normal group,a model group,and a Xiaozhongzhitong mixture group(Xiaozhong group),with 10 rats in each group.After intraperitoneal anesthesia,the model group and Xiaozhong group were completely freed with a pedicled skin flap of about 5 cm*6 cm in size on the back to simulate FIRI;the Xiaozhong group was given Xiaozhongzhitong Mixture(1.8 mL·kg-1)by gavage,once a day,the normal group and the model group were given normal saline(1.8 mL·kg-1)by gavage,once a day,and the rats in the three groups were continuously gavaged for 7 days;after 7 days,the dorsal skin flaps of the rats were collected for TMT proteomics analysis,screen out the differential proteins,perform Gene Ontology(GO)enrichment analysis,pathway enrichment(KEGG)analysis,common differential protein analysis,and establish Protein-Protein Interaction(PPI)network analysis.Results In this experiment,a total of 5005 proteins were screened by protein quantification,of which 4996 proteins were identified,and 141 up-regulated and 132 down-regulated differentially expressed proteins were screened out between the detumescence group/model group.GO enrichment analysis found that the differentially expressed proteins were mainly involved in molecular mechanisms such as organism cell metabolism,occurrence and combination of cellular components,catabolism,organelle formation,mitochondria formation,intracellular substance catalysis,molecular binding,hydrolase activation,and nucleotide regulation.KEGG pathway analysis found that the differentially expressed proteins were mainly involved in signaling pathways such as organismal cell metabolism,ribosome transcription and translation,proteasome system,and cell cycle.PPI analysis found that the differentially expressed co-expressed proteins Mrpl32,Psma2,Mrpl34,Psma4,Psmb1,and Srp9 were located at key nodes of the interaction network.Conclusion The treatment of FIRI with Xiaozhongzhitong Mixture may be related to the mitochondrial ribosome-related signaling pathway and the 20S proteasome system signaling pathway,among which the differential proteins Mrpl32,Psma2,Mrpl34,Psma4 and Psmb1 may be the key targets of treatment.