Inhibition of the P53/P21 Pathway Attenuates the Effects of Senescent Nucleus Pulposus Cell‐Derived Exosomes on the Senescence of Nucleus Pulposus Cells

Autor: Jing Chen, Chang-Chun Chen, Liang Xie, Wen-Liang Wang, Chuan-Qiang Shao, Yan-Xiang Zhang
Jazyk: angličtina
Rok vydání: 2021
Předmět:
Zdroj: Orthopaedic Surgery, Vol 13, Iss 2, Pp 583-591 (2021)
Orthopaedic Surgery
ISSN: 1757-7853
1757-7861
Popis: Objective The purpose of this paper is to investigate the effects of senescent nucleus pulposus cell (NPC)‐derived exosomes (SNPC‐Exo) and the roles of the P53/P21 pathway on the senescence of NPC. Methods The senescent phenotypes of NPC were induced by interleukin‐1β treatment. SNPC‐Exo was extracted from the culture medium of senescent NPC and purified by differential centrifugation. The structure of SNPC‐Exo was identified by transmission electron microscopy and western blot analysis was used to determine the exosomal marker proteins CD63 and Tsg101. Western blot analysis was performed to determine the relative expression levels of P16, P21, and P53 in NPC. Senescence‐associated β‐galactosidase (SA‐β‐gal) staining was used to stain the senescent NPC and a phase contrast microscope was used to observe and count the SA‐β‐gal staining of NPC. The proliferation of SNPC‐Exo‐treated NPC was assessed using growth curve analysis and the colony formation assay. The cell cycle of SNPC‐Exo‐treated NPC was determined by flow cytometry. NPC were transfected with siRNA to knock down P53 and P21 expression. Results Interleukin‐1β‐treated NPC had a higher percentage of SA‐β‐gal positive cells (45%) than the control group (20%) and showed an increase in the relative expression of P16, P21, and P53 (P
This study investigated the effects of SNPC‐Exo on the senescence of NPC. SNPC‐Exo‐treated NPC showed a senescence‐related phenotype. The inhibition of the P53/P21 pathway attenuated the senescence of NPC induced by SNPC‐Exo.
Databáze: OpenAIRE
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