1,25(OH)2D3 Mitigates Oxidative Stress‐Induced Damage to Nucleus Pulposus‐Derived Mesenchymal Stem Cells through PI3K/Akt Pathway

Intervertebral disc degeneration (IVDD) is one of the main causes of low back pain. The local environment of the degenerated intervertebral disc (IVD) increases oxidative stress and apoptosis of endogenous nucleus pulposus-derived mesenchymal stem cells (NPMSCs) and weakens its ability of endogenous repair ability in degenerated IVDs. A suitable concentration of 1α,25-dihydroxyvitamin D₃ (1,25(OH)₂D₃) has been certified to reduce oxidative stress and cell apoptosis. The current study investigated the protective effect and potential mechanism of 1,25(OH)₂D₃ against oxidative stress-induced damage to NPMSCs. The present results showed that 1,25(OH)₂D₃ showed a significant protective effect on NPMSCs at a concentration of 10^-10 M for 24 h. Protective effects of 1,25(OH)₂D₃ were also exhibited against H₂O₂-induced NPMSC senescence, mitochondrial dysfunction, and reduced mitochondrial membrane potential. The Annexin V/PI apoptosis detection assay, TUNEL assay, immunofluorescence, western blot, and real-time quantitative polymerase chain reaction assay showed that pretreatment with 1,25(OH)₂D₃ could alleviate H₂O₂-induced NPMSC apoptosis, including the apoptosis rate and the expression of proapoptotic-related (Caspase-3 and Bax) and antiapoptotic-related (Bcl-2) proteins. The intracellular expression of p-Akt increased after pretreatment with 1,25(OH)₂D₃. However, these protective effects of 1,25(OH)₂D₃ were significantly decreased after the PI3K/Akt pathway was inhibited by the LY294002 treatment. In vivo, X-ray, MRI, and histological analyses showed that 1,25(OH)₂D₃ treatment relieved the degree of IVDD in Sprague-Dawley rat disc puncture models. In summary, 1,25(OH)₂D₃ efficiently attenuated oxidative stress-induced NPMSC apoptosis and mitochondrial dysfunction via PI3K/Akt pathway and is a promising candidate treatment for the repair of IVDD.