Sang Ping, Li Xuepeng, Wang Ziyu
Department of Spine Surgery, Jilin Provincial People's Hospital, No. 1183, Gongnong Road, Changchun, 130021, Jilin, China.
Appl Biochem Biotechnol. 2024 Nov;196(11):8043-8057. doi: 10.1007/s12010-024-04953-z. Epub 2024 Apr 27.
Intervertebral disc degeneration (IVDD) is the primary cause of low back pain. Stem cell transplantation may be a possible approach to promote IVDD. This study was aimed to investigate the role of bone mesenchymal stem cells (BMSCs) in IVDD and the molecular mechanism. Annulus fibrosus cells (AFCs) were treated with tert-butyl hydroperoxide (TBHP) to induce oxidative stress injury. AFC biological functions were analyzed using a lactate dehydrogenase kit, enzyme-linked immunosorbent assay, flow cytometry, and western blot. The molecular mechanisms of BMSC functions were assessed using quantitative real-time PCR, western blot, immunoprecipitation (IP), co-IP, GST pull-down, and cycloheximide treatment. Furthermore, the impacts of BMSCs in IVDD progression in vivo were evaluated by magnetic resonance imaging (MRI) and H&E analysis. BMSCs inhibited TBHP-induced inflammation and pyroptosis in AFCs. Knockdown of SIRT1 reversed the effects on inflammation and pyroptosis of BMSCs. Moreover, SIRT1 promoted the deacetylation of ASC rather than NLRP3. SIRT1 interacted with ASC to reduce its protein stability, thereby negatively regulating ASC protein levels. In addition, BMSCs alleviated LPS-induced IVDD based on matrix hydrogels. BMSCs inhibited oxidative stress-induced pyroptosis and inflammation in AFCs, thereby alleviating IVDD, suggesting that BMSCs may contribute to treating intervertebral disc generation.
椎间盘退变(IVDD)是腰痛的主要原因。干细胞移植可能是促进IVDD恢复的一种可行方法。本研究旨在探讨骨髓间充质干细胞(BMSCs)在IVDD中的作用及其分子机制。用叔丁基过氧化氢(TBHP)处理纤维环细胞(AFCs)以诱导氧化应激损伤。使用乳酸脱氢酶试剂盒、酶联免疫吸附测定、流式细胞术和蛋白质印迹法分析AFC的生物学功能。使用定量实时PCR、蛋白质印迹法、免疫沉淀(IP)、免疫共沉淀、谷胱甘肽S-转移酶拉下实验和环己酰亚胺处理评估BMSC功能的分子机制。此外,通过磁共振成像(MRI)和苏木精-伊红(H&E)分析评估BMSCs对体内IVDD进展的影响。BMSCs抑制TBHP诱导的AFCs炎症和焦亡。敲低SIRT1可逆转BMSCs对炎症和焦亡的影响。此外,SIRT1促进ASC的去乙酰化而非NLRP3的去乙酰化。SIRT1与ASC相互作用以降低其蛋白质稳定性,从而负向调节ASC蛋白水平。此外,基于基质水凝胶,BMSCs减轻了脂多糖诱导的IVDD。BMSCs抑制氧化应激诱导的AFCs焦亡和炎症,从而减轻IVDD,表明BMSCs可能有助于治疗椎间盘退变。