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- Title
Differences In Protein Profiling Between Degenerated And Herniated Disc - A Proteomic Analysis.
- Authors
Rajasekaran, S.; Tangavel, Chitraa; Nayagam, Sharon Miracle; Aiyer, Siddharth; Muthurajan, Raveendran; Dharmalingam, Kuppamuthu
- Abstract
Introduction: We report the first comparison of proteomic profile in the disc herniations and disc degeneration. Proteomics will assist in understanding the differential expressions of low abundant proteins (expressed at a magnitude of 10-15, which might serve as a potent biomarker for understanding disease progression at various stages. Materials and Methods: We performed a comparative proteomic profiling of intervertebral disc tissue between 15 cases of disc herniation (DH) undergoing microlumbar discectomy and 5 cases of severe disc degeneration (DD) treated with lumbar fusion. Total proteins were extracted from all these samples using two buffers; radioimmuno precipitation assay buffer (RIPA) and 2% Sodium dodecyl sulphate (SDS). The resultant soluble and insoluble proteins were were cleaned using methanol: chloroform. Around 100 μg of total proteins were pre-fractionated on SDS-PAGE (Polyacrylamide gel electrophoresis) and proteins were analysed LC-MS/MS (Liquid Chromatography- tandem mass spectrometry) shotgun proteomics. Protein identification was performed using Proteome discoverer (PD) version 1.4.1.14 and Gene Ontology (GO) analysis for biological process, molecular function and cellular component of disc tissue was identified based on PD. Functional protein association network in the disc tissue were analysed using STRING database. Results: Total number of proteins identified in DH group was 945 and in DD group were 514. Pathways unique in DH are Vascular endothelial growth factor (VEGF) pathway, Transforming growth factor beta (TGF-β) signaling pathway, Platelet derived growth factor (PDGF) pathway, Epidermal growth factor (EGF) pathway, P-53 pathway, Wnt Signaling pathway, angiogenesis and apoptotic pathways. Presence of VEGF, TGF- β, PDGF pathway proteins such as Rho-GTPase activating 1 (VEGF), Inhibin beta A chain (TGF- β), PDGF factor subunit B, PDGF receptor like protein suggests angiogenesis. Similarly apoptotic pathway proteins Myosin 13 (Wnt signalling) and Heat shock protein family A5 supports IVD cells undergoing apoptosis during disc herniation. Interestingly analysis of the unique proteins of herniated discs using STRING protein functional association network shows around 106 proteins expressed in response to stress. Pathways unique to proteins to DD group are- axon mediated guidance pathway and vitamin D metabolic pathway. Protein semaphorin 3A (axon mediated guidance pathway) was observed in DD group which is a potent inhibitor of axon outgrowth and pathological innervations. Vitamin D binding protein levels were elevated in DD group. Analysis of the unique protein's functional association using STRING revealed, around 12 proteins specific to oxidative stress, 12 proteins of receptor mediated endocytosis and 25 proteins involved in vesicle mediated transport in DD group. Conclusion: This is the first comparison of proteomic profile in the disc herniations and disc degeneration. The disc herniation group showed a more abundance of proteins compared to disc degeneration group. This abundance was due to pro angiogenic, pro apoptotic and stress response proteins. In contrast disc degeneration group had more evidence of receptor mediated endocytosis and vesicle mediated transport proteins.
- Publication
Global Spine Journal, 2018, Vol 8, p272S
- ISSN
2192-5682
- Publication type
Article
- DOI
10.1177/2192568218771072