Research Paper Volume 14, Issue 10 pp 4572—4585
Lactotransferrin promotes intervertebral disc degeneration by regulating Fas and inhibiting human nucleus pulposus cell apoptosis
- 1 Department of Spine Surgery, Honghui Hospital, Xi'an Jiao tong University, Xi'an, Shaanxi 710000, PR China
- 2 Department of Spine Surgery, Lanzhou University Second Hospital, Lanzhou, Gansu 730000, PR China
Received: July 26, 2021 Accepted: April 11, 2022 Published: May 25, 2022
https://doi.org/10.18632/aging.204100How to Cite
Copyright: © 2022 Zhang et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Abstract
Background: In recent years, intervertebral disc (IVD) degeneration (IDD) has increased in age. There is still a lack of effective treatment in clinics, which cannot improve the condition of IDD at the level of etiology.
Objective: To explore IDD pathogenesis at the cellular and gene levels and investigate lactotransferrin (LTF) expression in IDD patients and its possible mechanism.
Methods: We downloaded the IDD data set from the Gene Expression Omnibus (GEO) database, screened the differentially expressed genes (DEGs) and hub genes and performed Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis to construct a protein–protein interaction (PPI) network. Subsequently, we verified LTF's regulatory mechanism through cell experiments. IL-1β was used to intervene in nucleus pulposus cells (NPCs) to construct the IDD cell model, and LTF and Fas expression was detected by qRT–PCR. LTF inhibitor, Fas inhibitor, LTF mimic, and Fas mimic were used to intervene in each group. Western blotting was used to detect Fas, Caspase-3, Bax, and Bcl-2 expression.
Results: A total of 131 DEGs and 10 hub genes were screened. LTF mRNA in the IDD model was significantly higher than that in the control group, while Fas' mRNA was significantly lower. When LTF was upregulated or downregulated in NPCs, apoptosis marker expression showed the opposite trend. The rescue test showed that LTF and Fas' overexpression greatly enhanced NPC apoptosis.
Conclusion: LTF promotes IDD progression by regulating Fas in NPCs, and it may be an effective gene therapy target.
Abbreviations
IVD: Intervertebral disc; IDD: Intervertebral disc degeneration; GEO: Gene expression omnibus; DEGs: Differentially expressed genes; KEGG: Kyoto encyclopedia of genes and genomes; PPI: Protein–protein interaction network; LTF: Lactotransferrin; NP: Nucleus pulposus; NPCs: Nucleus pulposus cells; ECM: Extracellular matrix; GSVA: Gene set variation analysis; SDS–PAGE: Sodium dodecyl sulfate–polyacrylamide gel electrophoresis; PVDF: Polyvinylidene fluoride.