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Abstract：
The aim of this study is to investigate the effects of POSTN on IL-1β induced inflammation, apoptosis, NF-κB pathway and intervertebral disc degeneration (IVDD) in Nucleus pulposus (NP) cells (NPCs). NP tissue samples with different Pfirrmann grades were collected from patients with different degrees of IVDD. Western blot and immunohistochemical staining were used to compare the expression of POSTN protein in NP tissues. Using the IL-1β-induced IVDD model, NPCs were transfected with lentivirus-coated si-POSTN to down-regulate the expression of POSTN and treated with CU-T12-9 to evaluate the involvement of NF-κB pathway. Western blot, immunofluorescence, and TUNEL staining were used to detect the expression changes of inflammation, apoptosis and NF-κB pathway-related proteins in NPCs. To investigate the role of POSTN in vivo, a rat IVDD model was established by needle puncture of the intervertebral disc. Rats were injected with lentivirus-coated si-POSTN, and H&E staining and immunohistochemical staining were performed. POSTN expression is positively correlated with the severity of IVDD in human. POSTN expression was significantly increased in the IL-1β-induced NPCs degeneration model. Downregulation of POSTN protects NPCs from IL-1β-induced inflammation and apoptosis. CU-T12-9 treatment reversed the protective effect of si-POSTN on NPCs. Furthermore, lentivirus-coated si-POSTN injection partially reversed NP tissue damage in the IVDD model in vivo. POSTN knockdown reduces inflammation and apoptosis of NPCs by inhibiting NF-κB pathway, and ultimately prevents IVDD. Therefore, POSTN may be an effective target for the treatment of IVDD.
摘要：
本研究旨在探讨POSTN对IL-1β诱导的炎症反应的影响,凋亡，NF-κB通路与髓核(NP)细胞(NPCs)椎间盘退变(IVDD)的关系.从具有不同IVDD程度的患者中收集具有不同Pfirrmann等级的NP组织样本。Westernblot和免疫组化染色比较POSTN蛋白在NP组织中的表达。使用IL-1β诱导的IVDD模型，用慢病毒包被的si-POSTN转染NPC以下调POSTN的表达，并用CU-T12-9处理以评估NF-κB途径的参与。蛋白质印迹,免疫荧光，和TUNEL染色检测炎症的表达变化,NPCs细胞凋亡和NF-κB通路相关蛋白。为了研究POSTN在体内的作用，通过椎间盘穿刺建立大鼠IVDD模型。大鼠注射慢病毒包被的si-POSTN，进行H&E染色和免疫组织化学染色。POSTN表达与人IVDD的严重程度呈正相关。POSTN表达在IL-1β诱导的NPCs变性模型中显著增加。POSTN的下调保护NPCs免受IL-1β诱导的炎症和凋亡。CU-T12-9处理逆转了si-POSTN对NPCs的保护作用。此外，慢病毒包被的si-POSTN注射液在体内部分逆转了IVDD模型中的NP组织损伤。POSTN敲低通过抑制NF-κB通路减少NPCs的炎症和凋亡,并最终阻止IVDD。因此,POSTN可能是治疗IVDD的有效靶点。
