N-Cadherin Attenuates High Glucose-Induced Nucleus Pulposus Cell Senescence Through Regulation of the ROS/NF-κB Pathway
N-Cadherin Attenuates High Glucose-Induced Nucleus Pulposus Cell Senescence Through Regulation of the ROS/NF-κB Pathway
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DOI：
10.1159/000489804
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发表时间：
2018-01-01
影响因子：
--
通讯作者：
Xu, Yichun
中科院分区：
文献类型：
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作者：
Hou, Gang;Zhao, Huiqing;Xu, Yichun
背景/目的：糖尿病（DM）是椎间盘退变的潜在病因。N-钙粘蛋白（N-CDH）有助于维持髓核（NP）细胞的细胞活力、细胞表型和基质生物合成。本研究旨在探讨N-CDH是否能减轻高糖诱导的NP细胞衰老及其可能的机制。方法：将大鼠NP细胞分别培养在基础培养基和含0.2M葡萄糖的基础培养基中。重组慢病毒载体用于增强NP细胞中的N-CDH表达。通过SA-β-Gal染色测量衰老相关的β-半乳糖苷酶（SA-β-Gal）活性。CCK-8法检测NP细胞增殖;端粒酶活性和细胞内活性氧（ROS）含量的检测采用特定的化学试剂盒，根据制造商的说明。流式细胞术检测G 0/G1期细胞阻滞。实时荧光定量PCR和Western blotting分析衰老标志物（p16和p53）和基质大分子（aggrecan和collagen II）的mRNA和蛋白表达。此外，还通过蛋白质印迹法分析p-NF-κ B表达以评价NF-κ B途径活性。结果如下：与对照组相比，高糖显著降低N-CDH表达，增加ROS产生和NF-κ B通路活性，并促进NP细胞衰老，这反映在SA-β-Gal活性和衰老标志物（p16和p53）表达的增加。高糖降低端粒酶活性和细胞增殖能力。然而，在高糖条件下，N-CDH过表达部分地减轻了NP细胞的衰老，降低了ROS含量，并抑制了NF-κ B通路的激活。结论：高糖降低N-CDH表达，促进NP细胞衰老。N-CDH过表达可通过调节ROS/NF-κ B B通路减轻高糖诱导的NP细胞衰老。这项研究表明，N-CDH是一个潜在的治疗目标，以减缓DM介导的椎间盘NP退变。(c)2018作者（S）由S发布。Karger AG，巴塞尔。
Background/Aims: Diabetes mellitus (DM) is a potential etiology of disc degeneration. N-cadherin (N-CDH) helps maintain the cell viability, cell phenotype and matrix biosynthesis of nucleus pulposus (NP) cells. Here, we mainly aimed to investigate whether N-CDH can attenuate high glucose-induced NP cell senescence and its potential mechanism. Methods: Rat NP cells were cultured in a base culture medium and base culture medium with a 0.2 M glucose concentration. Recombinant lentiviral vectors were used to enhance N-CDH expression in NP cells. Senescence-associated beta-galactosidase (SA-beta-Gal) activity was measured by SA-beta-Gal staining. NP cell proliferation was evaluated by CCK-8 assay. Telomerase activity and intracellular reactive oxygen species (ROS) content were tested by specific chemical kits according to the manufacturer's instructions. G0/G1 cell cycle arrest was evaluated by flow cytometry. Real-time PCR and Western blotting were used to analyze mRNA and protein expressions of senescence markers (p16 and p53) and matrix macromolecules (aggrecan and collagen II). Additionally, p-NF-kappa B expression was also analyzed by Western blotting to evaluate NF-kappa B pathway activity. Results: High glucose significantly decreased N-CDH expression, increased ROS generation and NF-kappa B pathway activity, and promoted NP cell senescence, which was reflected in the increase in SA-beta-Gal activity and senescence marker (p16 and p53) expression, compared to the control group. High glucose decreased telomerase activity and cell proliferation potency. However, N-CDH overexpression partially attenuated NP cell senescence, decreased ROS content and inhibited the activation of the NF-kappa B pathway under the high glucose condition. Conclusion: High glucose decreases N-CDH expression and promotes NP cell senescence. N-CDH overexpression can attenuate high glucose-induced NP cell senescence through the regulation of the ROS/NF-kappa B pathway. This study suggests that N-CDH is a potential therapeutic target to slow DM-mediated disc NP degeneration. (c) 2018 The Author(s) Published by S. Karger AG, Basel.