中国组织工程研究

中国组织工程研究 ›› 2026, Vol. 30 ›› Issue (31): 8190-8196.doi: 10.12307/2026.854

• 干细胞培养与分化 stem cell culture and differentiation • 上一篇    下一篇

胞苷/尿苷单磷酸激酶2调控脂多糖诱导人牙龈成纤维细胞的铁死亡

何家琪1，2，沈恒宇1，2，胥铭章1，2，3，严  欣1，2，张瑞烽1，2，余  科1，2，3   

  1. 西南医科大学附属口腔医院，1口腔颌面修复与再生泸州重点实验室，3种植科，四川省泸州市  646000；2西南医科大学口腔医学研究所，四川省泸州市  646000
  • 收稿日期:2025-10-17 接受日期:2026-01-22 出版日期:2026-11-08 发布日期:2026-05-25
  • 通讯作者: 余科，博士，副教授，西南医科大学附属口腔医院，口腔颌面修复与再生泸州重点实验室，种植科，四川省泸州市 646000；西南医科大学口腔医学研究所，四川省泸州市 646000
  • 作者简介:何家琪，男，2002年生，四川省眉山市人，汉族，西南医科大学在读本科，主要从事口腔种植临床与基础研究。
  • 基金资助:
    四川省医学会医学科研项目(S2024053)，项目负责人：余科

Cytidine/uridine monophosphate kinase 2 regulates ferroptosis of human gingival fibroblasts induced by lipopolysaccharide

He Jiaqi1, 2, Shen Hengyu1, 2, Xu Mingzhang1, 2, 3, Yan Xin1, 2, Zhang Ruifeng1, 2, Yu Ke1, 2, 3   

  1. 1Luzhou Key Laboratory of Oral & Maxillofacial Reconstruction and Regeneration, Affiliated Stomatological Hospital, Southwest Medical University, Luzhou 646000, Sichuan Province, China; 2Institute of Stomatology, Southwest Medical University, Luzhou 646000, Sichuan Province, China; 3Department of Implantology, Affiliated Stomatological Hospital, Southwest Medical University, Luzhou 646000, Sichuan Province, China
  • Received:2025-10-17 Accepted:2026-01-22 Online:2026-11-08 Published:2026-05-25
  • Contact: Yu Ke, MD, Associate professor, Luzhou Key Laboratory of Oral & Maxillofacial Reconstruction and Regeneration, Affiliated Stomatological Hospital, Southwest Medical University, Luzhou 646000, Sichuan Province, China; Institute of Stomatology, Southwest Medical University, Luzhou 646000, Sichuan Province, China; Department of Implantology, Affiliated Stomatological Hospital, Southwest Medical University, Luzhou 646000, Sichuan Province, China
  • About author:He Jiaqi, Luzhou Key Laboratory of Oral & Maxillofacial Reconstruction and Regeneration, Affiliated Stomatological Hospital, Southwest Medical University, Luzhou 646000, Sichuan Province, China; Institute of Stomatology, Southwest Medical University, Luzhou 646000, Sichuan Province, China
  • Supported by:
    Medical Research Project of Sichuan Medical Association, No. S2024053 (to YK)

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摘要：

文题释义：

铁死亡：是一种铁依赖性的新型细胞程序性死亡方式，由DIXON等人于2012年首次提出，这种死亡方式与铁代谢异常和脂质过氧化密切相关，作用分子、效应形态及生化特征均不同于凋亡、自噬、坏死和焦亡等其他细胞死亡形式。
胞苷/尿苷单磷酸激酶2：是一种定位于线粒体的激酶，负责催化胞苷单磷酸和尿苷单磷酸磷酸化为相应的二磷酸形式，为核苷酸合成提供前体物质，参与维持线粒体DNA的复制与修复，影响能量代谢稳态，已被证实在抵抗细菌、病毒感染和抗肿瘤治疗中具有重要作用。

摘要
背景：铁死亡是一种铁依赖性细胞程序性死亡，与牙周、种植体周炎症性疾病关系密切，胞苷/尿苷单磷酸激酶2是一种定位于线粒体参与调控组织细胞活化和炎症反应的激酶，二者在种植体周炎中的调控机制尚不清楚。
目的：通过脂多糖构建种植体周炎的体外细胞模型，探讨胞苷/尿苷单磷酸激酶2对脂多糖诱导的人牙龈成纤维细胞铁死亡的调控机制。
方法：①采用组织块培养法培养人牙龈成纤维细胞，分别加入0(正常对照组)、10 μg/mL脂多糖刺激12 h，通过RT-qPCR和Western blot检测铁死亡相关标志物谷胱甘肽过氧化物酶4、溶质载体家族7成员11和酰基辅酶A合成酶长链家族成员4及胞苷/尿苷单磷酸激酶2的mRNA与蛋白表达，通过活性氧染色及C11-Bodipy染色检测脂质过氧化水平，通过比色法检测细胞内丙二醛、超氧化物歧化酶水平以及还原型谷胱甘肽/氧化型谷胱甘肽比率；②siRNA沉默胞苷/尿苷单磷酸激酶2，再以10 μg/mL脂多糖刺激细胞12 h，检测上述指标的改变。
结果与结论：①10 μg/mL脂多糖刺激人牙龈成纤维细胞12 h后，铁死亡标志物酰基辅酶A合成酶长链家族成员4表达上调，铁死亡负性调控因子谷胱甘肽过氧化物酶4和溶质载体家族7成员11表达下调，活性氧释放增加，丙二醛水平升高，超氧化物歧化酶活性及还原型谷胱甘肽/氧化型谷胱甘肽比率降低，胞苷/尿苷单磷酸激酶2表达上调；②siRNA沉默胞苷/尿苷单磷酸激酶2后，再以10 μg/mL脂多糖刺激细胞12 h，酰基辅酶A合成酶长链家族成员4表达下调，谷胱甘肽过氧化物酶4和溶质载体家族7成员11表达上调，活性氧释放减少，丙二醛水平降低，超氧化物歧化酶活性及还原型谷胱甘肽/氧化型谷胱甘肽比率升高。结果表明，胞苷/尿苷单磷酸激酶2是种植体周炎牙龈成纤维细胞铁死亡的关键调控基因，沉默胞苷/尿苷单磷酸激酶2可抑制脂多糖诱导的人牙龈成纤维细胞铁死亡。

关键词: 胞苷/尿苷单磷酸激酶2, 铁死亡, 脂多糖, 人牙龈成纤维细胞, 种植体周炎, 谷胱甘肽过氧化物酶4, 活性氧, 超氧化物歧化酶

Abstract: BACKGROUND: Ferroptosis is an iron-dependent form of programmed cell death closely associated with periodontal and peri-implant inflammatory diseases. Cytidine/uridine monophosphate kinase 2 is a kinase localized in mitochondria that participates in regulating tissue cell activation and inflammatory responses. The regulatory mechanisms of both in peri-implantitis remain unclear.
OBJECTIVE: To construct an in vitro cell model of peri-implantitis using lipopolysaccharide and explore the regulatory mechanism of cytidine/uridine monophosphate kinase 2 on lipopolysaccharide-induced ferroptosis in human gingival fibroblasts.
METHODS: (1) Human gingival fibroblasts were cultured using the tissue explant culture method and treated with 0 μg/mL (normal control group) or 10 μg/mL lipopolysaccharide for 12 hours. The mRNA and protein expression levels of ferroptosis-related markers glutathione peroxidase 4, solute carrier family 7 member 11, and acyl-CoA synthetase long-chain family member 4, as well as cytidine/uridine monophosphate kinase 2, were detected by RT-qPCR and western blot assay. Lipid peroxidation levels were detected by reactive oxygen species staining and C11-Bodipy staining. Intracellular malondialdehyde, superoxide dismutase levels, and the reduced glutathione/oxidized glutathione ratio were detected by colorimetric methods. (2) Cytidine/uridine monophosphate kinase 2 was silenced using siRNA, and then cells were stimulated with 10 μg/mL lipopolysaccharide for 12 hours. The changes in the above indicators were then detected.
RESULTS AND CONCLUSION: (1) After stimulating human gingival fibroblasts with 10 μg/mL lipopolysaccharide for 12 hours, the expression of the ferroptosis marker acyl-CoA synthetase long-chain family member 4 was upregulated, while the expression of the negative ferroptosis regulators glutathione peroxidase 4 and solute carrier family 7 member 11 was downregulated. Reactive oxygen species release increased, malondialdehyde levels increased, superoxide dismutase activity and the reduced glutathione/oxidized glutathione ratio decreased, and cytidine/uridine monophosphate kinase 2 expression was upregulated. (2) After silencing cytidine/uridine monophosphate kinase 2 with siRNA, and then stimulating the cells with 10 μg/mL lipopolysaccharide for 12 hours, acyl-CoA synthetase long-chain family member 4 expression was downregulated, glutathione peroxidase 4 and solute carrier family 7 member 11 expression were upregulated, reactive oxygen species release decreased, malondialdehyde levels decreased, and superoxide dismutase activity and the reduced glutathione/oxidized glutathione ratio increased. These results indicate that cytidine/uridine monophosphate kinase 2 is a key regulatory gene for ferroptosis in gingival fibroblasts during peri-implantitis, and silencing cytidine/uridine monophosphate kinase 2 can inhibit lipopolysaccharide-induced ferroptosis in human gingival fibroblasts.

Key words: cytidine/uridine monophosphate kinase 2, ferroptosis, lipopolysaccharide, human gingival fibroblasts, peri-implantitis, glutathione peroxidase 4, reactive oxygen species, superoxide dismutase

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