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轴向压应力过载激活了伴有椎间盘退变的成人退变性脊柱侧凸的髓核细胞中的 Wnt/β-连环蛋白通路。

Overloaded axial stress activates the Wnt/β-Catenin pathway in nucleus pulposus cells of adult degenerative scoliosis combined with intervertebral disc degeneration.

机构信息

Department of Orthopedics, The People's Liberation Army Joint Logistic Support Force 920th Hospital, No. 212 Daguan Rd, Kunming, Yunnan, 650032, China.

出版信息

Mol Biol Rep. 2023 Jun;50(6):4791-4798. doi: 10.1007/s11033-023-08390-9. Epub 2023 Apr 8.

DOI:10.1007/s11033-023-08390-9
PMID:37031322
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10209306/
Abstract

BACKGROUND

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.

METHODS

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).

RESULTS

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.

CONCLUSIONS

Overloaded axial pressure activates the Wnt/β-Catenin pathway to suppress proliferation and facilitate apoptosis of NPC in ADS-IVDD patients.

摘要

背景

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。

方法

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。

结果

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。

结论

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。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3c42/10209306/8d1b1aa37669/11033_2023_8390_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3c42/10209306/1660962d763c/11033_2023_8390_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3c42/10209306/e9275106911f/11033_2023_8390_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3c42/10209306/04b6734488d9/11033_2023_8390_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3c42/10209306/8d1b1aa37669/11033_2023_8390_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3c42/10209306/1660962d763c/11033_2023_8390_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3c42/10209306/e9275106911f/11033_2023_8390_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3c42/10209306/04b6734488d9/11033_2023_8390_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3c42/10209306/8d1b1aa37669/11033_2023_8390_Fig4_HTML.jpg

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