中国组织工程研究

中国组织工程研究 ›› 2026, Vol. 30 ›› Issue (29): 7520-7528.doi: 10.12307/2026.216

• 骨组织构建 bone tissue construction • 上一篇    下一篇

调控髓核细胞线粒体动力学平衡抑制髓核细胞凋亡

张治龙，王海英，马风华，侯彦杰   

  1. 新疆医科大学第二附属医院脊柱外科，新疆维吾尔自治区乌鲁木齐市   830000 


  • 收稿日期:2025-11-25 修回日期:2025-12-23 出版日期:2026-10-18 发布日期:2026-03-03
  • 通讯作者: 侯彦杰，硕士，主任医师，新疆医科大学第二附属医院脊柱外科，新疆维吾尔自治区乌鲁木齐市 830000
  • 作者简介:张治龙，男，1986年生，河南省兰考县人，汉族，2014年新疆医科大学毕业，硕士，主治医师，主要从事脊柱及关节退行性疾患的研究。
  • 基金资助:
    新疆天山英才医药卫生高层次人才培养计划项目(TSYC202301B124)，项目负责人：侯彦杰

Regulating mitochondrial dynamics balance in nucleus pulposus cells inhibits cell apoptosis

Zhang Zhilong, Wang Haiying, Ma Fenghua, Hou Yanjie   

  1. Department of Spine Surgery, The Second Affiliated Hospital of Xinjiang Medical University, Urumqi 830000, Xinjiang Uygur Autonomous Region, China 
  • Received:2025-11-25 Revised:2025-12-23 Online:2026-10-18 Published:2026-03-03
  • Contact: Hou Yanjie, MD, Chief physician, Department of Spine Surgery, The Second Affiliated Hospital of Xinjiang Medical University, Urumqi 830000, Xinjiang Uygur Autonomous Region, China
  • About author:Zhang Zhilong, MS, Department of Spine Surgery, The Second Affiliated Hospital of Xinjiang Medical University, Urumqi 830000, Xinjiang Uygur Autonomous Region, China
  • Supported by:
    Xinjiang Tianshan Elite Medical and Health High-Level Talent Cultivation Program Project, No. TSYC202301B124 (to HYJ)

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摘要：



文题释义：
髓核细胞退变：指髓核细胞在生理或病理因素影响下，出现功能性细胞数量减少、细胞形态和结构异常、生物合成能力下降等一系列变化的过程。
线粒体动力学平衡：指线粒体在细胞内通过持续的融合与裂变过程，维持其形态、数量、分布及功能状态的动态稳定，这一平衡对细胞的能量供应、代谢调控、细胞周期进程以及细胞命运决定(如凋亡、分化或坏死)等生命活动至关重要。

背景：线粒体功能障碍在椎间盘退变进程中的核心作用逐渐被认知，其中Sirt3作为线粒体主要去乙酰化酶，可介导AMPK通路激活，不仅能直接磷酸化抑制Drp1活性，还可通过调控下游信号间接影响线粒体功能。但Sirt3与AMPK/Drp1通路在椎间盘退变髓核细胞中的具体作用机制尚不明确。
目的：探究Sirt3是否通过介导AMPK/Drp1通路调控叔丁基过氧化氢诱导的髓核细胞线粒体动力学平衡，抑制髓核细胞凋亡。
方法：体外培养人髓核细胞，通过叔丁基过氧化氢氧化损伤建立髓核细胞退变模型。将人髓核细胞分为以下几组：正常组、模型组、模型+oe-NC组、模型+oe-Sirt3组、模型+oe-Sirt3+Compound C(AMPK抑制剂)组、Compound C组，各组分别处理24 h。通过CCK-8检测细胞活力，流式细胞术检测细胞凋亡，western blot检测细胞凋亡相关蛋白(Bax和Bcl2)、椎间盘退变相关蛋白(聚集蛋白聚糖、Ⅱ型胶原)及Sirt3的表达，并检测线粒体分裂相关蛋白(Fis1、Mff)和线粒体融合相关蛋白(Mfn1、Mfn2)及AMPK/Drp1信号通路相关蛋白的表达水平。试剂盒检测三磷酸腺苷和活性氧含量，RT-qPCR检测线粒体mtDNA拷贝数。
结果与结论：①与正常组相比，模型组细胞活力及Bcl2、聚集蛋白聚糖、Ⅱ型胶原、Sirt3表达显著降低，细胞凋亡率及Bax水平显著升高(均P < 0.05)，说明髓核细胞退变模型建立成功。②与正常组相比，模型组三磷酸腺苷、线粒体膜电位及mtDNA拷贝数、Mfn1、Mfn2表达显著降低，活性氧、Fis1、Mff水平升高(均P < 0.05)，说明髓核细胞退变模型线粒体动力学失衡；而在模型+oe-Sirt3组中过表达Sirt3能抑制叔丁基过氧化氢诱导的椎间盘髓核细胞凋亡，提高细胞活力，改善线粒体动力学平衡，促进AMPK/Drp1通路激活，抑制线粒体分裂，从而抑制叔丁基过氧化氢诱导的髓核细胞凋亡；模型+oe-Sirt3+Compound C组抑制AMPK/Drp1信号部分逆转过表达Sirt3对叔丁基过氧化氢诱导的椎间盘髓核细胞凋亡和线粒体功能动力学平衡的改善作用。结果提示Sirt3是髓核细胞凋亡的潜在靶点，可作为椎间盘退变治疗的新方向。

https://orcid.org/0009-0008-9812-5165 (张治龙) 


中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

关键词: 髓核细胞, 线粒体动力学, 叔丁基过氧化氢, 椎间盘退变, 细胞凋亡

Abstract: BACKGROUND: Mitochondrial dysfunction is increasingly recognized as a key factor during intervertebral disc degeneration. Sirt3, a major mitochondrial deacetylase, mediates AMPK pathway activation by directly phosphorylating and inhibiting Drp1 activity while indirectly regulating mitochondrial function through downstream signaling. However, the specific mechanisms of Sirt3 and the AMPK/Drp1 pathway in nucleus pulposus cells during intervertebral disc degeneration remain unclear.
OBJECTIVE: To investigate whether Sirt3 regulates mitochondrial dynamics balance in nucleus pulposus cells induced by tert-butyl hydroperoxide by mediating the AMPK/Drp1 pathway, thereby inhibiting cell apoptosis.
METHODS: Human nucleus pulposus cells were cultured in vitro, and a degeneration model was established by oxidative damage with tert-butyl hydroperoxide. Cells were divided into the following groups: control, model, model + oe-NC, model + oe-Sirt3, model + oe-Sirt3 + Compound C (AMPK inhibitor), and Compound C. Each group was treated for 24 hours. Cell viability was assessed using the Cell Counting Kit-8 assay, apoptosis was detected by flow cytometry, and the expression levels of apoptosis-related proteins (Bax and Bcl2), disc degeneration-related proteins (aggrecan and type II collagen), Sirt3, mitochondrial fission-related proteins (Fis1 and Mff), mitochondrial fusion-related proteins (Mfn1 and Mfn2), and AMPK/Drp1 signaling pathway-related proteins were determined using western blot analysis. Adenosine triphosphate and reactive oxygen species levels were measured using kits, while the number of mitochondrial mtDNA copies was assessed by quantitative reverse transcription-polymerase chain reaction.
RESULTS AND CONCLUSION: (1) Compared with the control group, cell viability and expression of Bcl2, aggrecan, type II collagen, and Sirt3 were significantly decreased, while apoptosis rate and Bax expression levels significantly increased in the model group (all P < 0.05), confirming successful establishment of the nucleus pulposus cell degeneration model. (2) Compared with the control group, ATP, mitochondrial membrane potential, mtDNA copy number, and Mfn1/Mfn2 expression were significantly decreased, while reactive oxygen species, Fis1, and Mff levels were significantly increased (all P < 0.05) in the model group, indicating mitochondrial dysregulation in the nucleus pulposus cell degeneration model. In the model + oe-Sirt3 group, Sirt3 overexpression inhibited tert-butyl hydroperoxide-induced nucleus pulposus cell apoptosis, enhanced cell viability, restored mitochondrial dynamics balance, promoted AMPK/Drp1 pathway activation, and inhibited mitochondrial fission, thereby suppressing tert-butyl hydroperoxide-induced nucleus pulposus cell apoptosis. Inhibition of AMPK/Drp1 signaling partially reversed the beneficial effects of Sirt3 overexpression on tert-butyl hydroperoxide-induced nucleus pulposus cell apoptosis and mitochondrial dynamics balance in the model + oe-Sirt3 + Compound C group. These findings suggest Sirt3 as a potential therapeutic target for nucleus pulposus cell apoptosis, offering a novel therapeutic approach for intervertebral disc degeneration. 

Key words: nucleus pulposus cells, mitochondrial dynamics, tert-butyl hydroperoxide, intervertebral disc degeneration, cell apoptosis

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