Journal of Pain Research (Mar 2024)
Tandem Mass Tag-Based Proteomic Analysis of Normal and Degenerated Human Intervertebral Discs
Abstract
Yang Fu,* Xiao-Qin Huang,* Hang-Bo Qu, Yan Zhi Ge, Xuan-Liang Ru Department of Orthopedics, Zhejiang Hospital, Hangzhou, Zhejiang Province, People’s Republic of China*These authors contributed equally to this workCorrespondence: Xuan-Liang Ru, Department of Orthopedics, Zhejiang Hospital, No. 1229 Gudun Road, Hangzhou, Zhejiang Province, 310013, People’s Republic of China, Tel +86-133-3612-4875, Email [email protected]: Intervertebral disc degeneration (IVDD) is the main cause of low back pain (LBP), but the specific regulatory factors, pathways and specific molecular mechanisms remain unclear.Methods: We identified and quantitatively analyzed Pfirrmann Grade II (n=3) and Pfirrmann Grade IV (n=3) pulposus samples via MRI. The differential abundance of proteins in the samples was determined and quantitatively analyzed by relative and absolute quantitative analysis of the isotope marker levels combined with the liquid chromatography–tandem mass spectrometry (LC‒MSMS/MS).Results: A total of 70 proteins (30 significantly increased proteins (> 1.2-fold change) and 40 significantly decreased proteins (< 0.8-fold change)) showed different levels among the groups. Kyoto Encyclopedia of Genes and Genomes and Gene Ontology (GO) enrichment analyses and Western blot analysis showed that CYCS, RAC1, and PSMD14 may play important roles in IVDD and that Epstein‒Barr virus infection, viral myocarditis, colorectal cancer, nonalcoholic fatty liver disease (NAFLD) and amyotrophic lateral sclerosis (ALS) are the main pathways involved in IVDD.Conclusion: CYCS, RAC1 and PSMD14 may play important roles in IVDD, and Epstein‒Barr virus infection, viral myocarditis, colorectal cancer, NAFLD and ALS may be the main pathways involved in IVDD.Keywords: TMT, LC‒MS/MS, proteomics, low back pain, nucleus pulposus