Department of Orthopedics, The People's Liberation Army Joint Logistic Support Force 920th Hospital, No. 212 Daguan Rd, Kunming, Yunnan, 650032, China.
Mol Biol Rep. 2023 Jun;50(6):4791-4798. doi: 10.1007/s11033-023-08390-9. Epub 2023 Apr 8.
Intervertebral disc degeneration (IVDD) is the initiating factor of adult degenerative scoliosis (ADS), and ADS further accelerates IVDD, creating a vicious cycle. Nevertheless, the role of the Wnt/β-Catenin pathway in ADS combined with IVDD (ADS-IVDD) remains a mystery. Accordingly, this study was proposed to investigate the effect of axial stress on the Wnt/β-Catenin pathway in nucleus pulposus cells (NPCs) isolated from DS-IVDD patients.
Normal NPCs (N-NPC) were purchased and the NPCs of young (25-30 years; Y-NPC) and old (65-70 years; O-NPC) from ADS-IVDD patients were primary cultured. After treatment of NPC with overloaded axial pressure, CCK-8 and Annexin V-FITC kits were applied for detecting proliferation and apoptosis of N-NPC, Y-NPC and O-NPC, and western blotting was performed to assess the expression of Wnt 3a, β-Catenin, NPC markers and apoptosis markers (Bax, Bcl2 and Caspase 3).
N-NPC, Y-NPC and O-NPC were mainly oval, polygonal and spindle-shaped with pseudopods, and the cell morphology tended to be flattened with age. N-NPC, Y-NPC and O-NPC were capable of synthesizing proteoglycans and expressing the NPC markers (Collagen II and Aggrecan). Notably, the expression of Wnt 3a, β-Catenin, Collagen II and Aggrecan was reduced in N-NPC, Y-NPC and O-NPC in that order. After overload axial stress treatment, cell viability of N-NPC and Y-NPC was significantly reduced, and the percentage of apoptosis and expression of Wnt 3a and β-Catenin were significantly increased.
Overloaded axial pressure activates the Wnt/β-Catenin pathway to suppress proliferation and facilitate apoptosis of NPC in ADS-IVDD patients.
椎间盘退变(IVDD)是成人退变性脊柱侧凸(ADS)的起始因素,而 ADS 进一步加速 IVDD,形成恶性循环。然而,Wnt/β-连环蛋白通路在 ADS 合并 IVDD(ADS-IVDD)中的作用仍不清楚。因此,本研究旨在探讨轴向压力对 ADS-IVDD 患者髓核细胞(NPC)中 Wnt/β-连环蛋白通路的影响。
购买正常 NPC(N-NPC),并对 ADS-IVDD 患者的年轻(25-30 岁;Y-NPC)和老年(65-70 岁;O-NPC)NPC 进行原代培养。用超负荷轴向压力处理 NPC 后,用 CCK-8 和 Annexin V-FITC 试剂盒检测 N-NPC、Y-NPC 和 O-NPC 的增殖和凋亡情况,用 Western blot 检测 Wnt 3a、β-连环蛋白、NPC 标志物和凋亡标志物(Bax、Bcl2 和 Caspase 3)的表达。
N-NPC、Y-NPC 和 O-NPC 主要呈椭圆形、多边形和梭形,有伪足,随着年龄的增长,细胞形态趋于扁平。N-NPC、Y-NPC 和 O-NPC 均能合成蛋白聚糖并表达 NPC 标志物(Collagen II 和 Aggrecan)。值得注意的是,Wnt 3a、β-连环蛋白、Collagen II 和 Aggrecan 的表达在 N-NPC、Y-NPC 和 O-NPC 中依次降低。在超负荷轴向压力处理后,N-NPC 和 Y-NPC 的细胞活力明显降低,凋亡百分比和 Wnt 3a、β-连环蛋白的表达明显增加。
超负荷轴向压力激活 Wnt/β-连环蛋白通路,抑制 ADS-IVDD 患者 NPC 的增殖,促进其凋亡。