Isobaric tag for relative and absolute quantitation-based proteomics for investigating the effect of Guasha on lumbar disc herniation in rats

Abstract

Objective: The objective of the study is to examine the possible mechanism by which Guasha (scraping therapy) affects the expression profiles of proteins in a lumbar disc herniation (LDH) rat model using isobaric tags for relative and absolute quantitation (iTRAQ)-based proteomics. Methods: Thirty-six rats were used in this study. LDH rats were subjected to noncompressive LDH surgeries. Rats were randomly divided into the model and Guasha groups. Guasha was applied on alternate days for a total of nine times (three courses). At the end of each course, six rats were randomly selected from each group and their blood samples were collected. iTRAQ labeling was used to examine the mechanism of action of Guasha against LDH. The molecular functions, cellular components, and biological processes were analyzed using gene ontology analysis. The Ingenuity Pathway Analysis database was used to identify canonical pathways involving these proteins. Results: Compared to the model group, 198, 182, and 170 proteins were identified as differentially expressed at the three respective Guasha treatment courses. Pathways, including focal adhesion kinase signaling, acute-phase response signaling, and the LXR/RXR activation pathway, were closely related to the effects of Guasha in LDH rats. Furthermore, Rac1, Orm1, and Hpx were validated by western blotting, and the results were consistent with the protein expression levels observed using the iTRAQ method. Conclusion: Guasha could not only regulate the pathological changes related to LDH, but also achieve therapeutic effects by stimulating physiological changes. Our results offer a better understanding of the effects of Guasha on LDH.