Toll-like receptor 2 induced senescence in intervertebral disc cells of patients with back pain can be attenuated by o-vanillin

Arthritis Research & Therapy

Oded Rabau¹  Hosni Cherif²  Peter Jarzem²  Matthew Mannarino²  Li Li²  Michael H. Weber²  Jean A. Ouellet³  Lisbet Haglund⁴  Kai Sheng⁵ 

[1] Department of Surgery, McGill Scoliosis and Spine Group, McGill University, Montreal, Canada;Shriner’s Hospital for Children, Montreal, Canada;Department of Surgery, Orthopaedic Research Lab, McGill University, Montreal, Canada;Department of Surgery, McGill Scoliosis and Spine Group, McGill University, Montreal, Canada;Department of Surgery, The Research Institute of McGill University Health Center, Montreal, Canada;Department of Surgery, Orthopaedic Research Lab, McGill University, Montreal, Canada;Department of Surgery, McGill Scoliosis and Spine Group, McGill University, Montreal, Canada;Department of Surgery, The Research Institute of McGill University Health Center, Montreal, Canada;Shriner’s Hospital for Children, Montreal, Canada;Department of Surgery, Orthopaedic Research Lab, McGill University, Montreal, Canada;Department of Surgery, McGill Scoliosis and Spine Group, McGill University, Montreal, Canada;Department of Surgery, The Research Institute of McGill University Health Center, Montreal, Canada;Shriner’s Hospital for Children, Montreal, Canada;Department of Surgery, Montreal General Hospital, McGill University Health Centre, Room C9.173,1650 Cedar Ave, H3G 1A4, Montreal, QC, Canada;Department of Surgery, Orthopaedic Research Lab, McGill University, Montreal, Canada;Department of Surgery, McGill Scoliosis and Spine Group, McGill University, Montreal, Canada;Shriner’s Hospital for Children, Montreal, Canada;

关键词: Intervertebral disc;    Senescence;    Toll-like receptor 2;    o-Vanillin;    Degeneration;    Inflammation;    Senolytics;    Back pain;   

DOI  :  10.1186/s13075-021-02504-z

来源: Springer

【 摘 要 】

BackgroundThere is an increased level of senescent cells and toll-like teceptor-1, -2, -4, and -6 (TLR) expression in degenerating intervertebral discs (IVDs) from back pain patients. However, it is currently not known if the increase in expression of TLRs is related to the senescent cells or if it is a more general increase on all cells. It is also not known if TLR activation in IVD cells will induce cell senescence.MethodsCells from non-degenerate human IVD were obtained from spine donors and cells from degenerate IVDs came from patients undergoing surgery for low back pain. Gene expression of TLR-1,2,4,6, senescence and senescence-associated secretory phenotype (SASP) markers was evaluated by RT-qPCR in isolated cells. Matrix synthesis was verified with safranin-O staining and Dimethyl-Methylene Blue Assay (DMMB) confirmed proteoglycan content. Protein expression of p16INK4a, SASP factors, and TLR-2 was evaluated by immunocytochemistry (ICC) and/or by enzyme-linked immunosorbent assay (ELISA).ResultsAn increase in senescent cells was found following 48-h induction with a TLR-2/6 agonist in cells from both non-degenerate and degenerating human IVDs. Higher levels of SASP factors, TLR-2 gene expression, and protein expression were found following 48-h induction with TLR-2/6 agonist. Treatment with o-vanillin reduced the number of senescent cells, and increased matrix synthesis in IVD cells from back pain patients. Treatment with o-vanillin after induction with TLR-2/6 agonist reduced gene and protein expression of SASP factors and TLR-2. Co-localized staining of p16INK4a and TLR-2 demonstrated that senescent cells have a high TLR-2 expression.ConclusionsTaken together our data demonstrate that activation of TLR-2/6 induce senescence and increase TLR-2 and SASP expression in cells from non-degenerate IVDs of organ donors without degeneration and back pain and in cells from degenerating human IVD of patients with disc degeneration and back pain. The senescent cells showed high TLR-2 expression suggesting a link between TLR activation and cell senescence in human IVD cells. The reduction in senescence, SASP, and TLR-2 expression suggest o-vanillin as a potential disease-modifying drug for patients with disc degeneration and back pain.

【 授权许可】

CC BY