中国组织工程研究

中国组织工程研究 ›› 2025, Vol. 29 ›› Issue (32): 6877-6884.doi: 10.12307/2025.921

• 口腔组织构建 oral tissue construction • 上一篇    下一篇

大鼠颞下颌关节骨关节炎模型中线粒体肌酸激酶2表达及在炎症进展中的作用

尼格阿依·艾合麦提1，2，伊丽丹娜·地里夏提1，2，安  玮1，2，买买提吐逊·吐尔地1，2   

  1. 1新疆医科大学第一附属医院(附属口腔医院)口腔颌面创伤正颌外科，新疆维吾尔自治区乌鲁木齐市  830054；2新疆维吾尔自治区口腔医学研究所，新疆维吾尔自治区乌鲁木齐市  830054
  • 收稿日期:2024-10-14 接受日期:2024-11-26 出版日期:2025-11-18 发布日期:2025-04-25
  • 通讯作者: 买买提吐逊·吐尔地，博士，主任医师，新疆医科大学第一附属医院(附属口腔医院)口腔颌面创伤正颌外科，新疆维吾尔自治区乌鲁木齐市 830054；新疆维吾尔自治区口腔医学研究所，新疆维吾尔自治区乌鲁木齐市 830054
  • 作者简介:尼格阿依·艾合麦提，女，1994年生，新疆维吾尔自治区乌鲁木齐市人，维吾尔族，在读硕士，主要从事颌面创伤外科研究。
  • 基金资助:
    国家自然科学基金项目(8236030005)，项目名称：线粒体内关键基因CKMT2的氧化修饰介导P53/MAPK信号通路促进颞下颌关节骨关节炎髁突软骨细胞凋亡的分子机制研究，项目负责人：买买提吐逊·吐尔地

Expression of mitochondrial creatine kinase 2 in a rat model of temporomandibular joint osteoarthritis and its role in inflammation progression

Nigeayi · Aihemaiti1, 2, Yilidanna · Dilixiati1, 2, An Wei1, 2, Maimaitituxun · Tuerdi1, 2   

  1. 1Department of Oral and Maxillofacial Trauma and Orthognathic Surgery, the First Affiliated Hospital (Affiliated Stomatological Hospital) of Xinjiang Medical University, Urumqi 830054, Xinjiang Uygur Autonomous Region, China; 2Xinjiang Uygur Autonomous Region Institute of Stomatology, Urumqi 830054, Xinjiang Uygur Autonomous Region, China
  • Received:2024-10-14 Accepted:2024-11-26 Online:2025-11-18 Published:2025-04-25
  • Contact: Maimaitituxun · Tuerdi, PhD, Chief physician, Department of Oral and Maxillofacial Trauma and Orthognathic Surgery, the First Affiliated Hospital (Affiliated Stomatological Hospital) of Xinjiang Medical University, Urumqi 830054, Xinjiang Uygur Autonomous Region, China; Xinjiang Uygur Autonomous Region Institute of Stomatology, Urumqi 830054, Xinjiang Uygur Autonomous Region, China
  • About author:Nigeayi · Aihemaiti, Master candidate, Department of Oral and Maxillofacial Trauma and Orthognathic Surgery, the First Affiliated Hospital (Affiliated Stomatological Hospital) of Xinjiang Medical University, Urumqi 830054, Xinjiang Uygur Autonomous Region, China; Xinjiang Uygur Autonomous Region Institute of Stomatology, Urumqi 830054, Xinjiang Uygur Autonomous Region, China
  • Supported by:
    National Natural Science Foundation of China, No. 8236030005 (to MT)

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摘要：


文题释义：
颞下颌关节骨关节炎：是一种病因复杂、发病机制不明的口腔颌面部常见疾病，属于颞下颌关节紊乱病的重症类型，病理特征是进行性软骨降解、软骨下骨重塑以及滑膜组织的慢性炎症。
线粒体肌酸激酶2：又称为肌节型肌酸激酶，是肌酸激酶同工酶之一。线粒体肌酸激酶2基因位于人类第5号染色体正链上，包含11个外显子，序列长度超过37 kb，在 ATP合成及细胞内能量转运过程中发挥重要作用。

背景：颞下颌关节骨关节炎的病因不明，尚无特效根治手段，治疗以缓解症状为主，因此，找到有效的早期分子诊断标志物或潜在的治疗靶点意义重大。
目的：探讨大鼠颞下颌关节骨关节炎中线粒体肌酸激酶2表达变化及其在炎症进展中的作用。
方法：①动物实验：采用随机数字表法将20只SD大鼠随机分为对照组和单侧前牙反牙合组(n=10)，单侧前牙反牙合组构建左侧颞下颌关节骨关节炎模型，造模4周后，通过苏木精-伊红染色、番红O-固绿染色观察下颌髁突软骨与软骨下骨病理变化，免疫组化染色、qRT-PCR检测白细胞介素1β、基质金属蛋白酶13、Ⅱ型胶原、蛋白聚糖、线粒体肌酸激酶2的蛋白与mRNA表达。②细胞实验：将第3代大鼠下颌髁突软骨细胞分2组培养，对照组常规培养，模型组加入白细胞介素1β处理诱导炎症细胞模型，白细胞介素1β处理24 h，Western blot检测基质金属蛋白酶13、Ⅱ型胶原蛋白表达；白细胞介素1β处理0，12，24，48，72 h，Western blot检测模型组细胞中线粒体肌酸激酶2蛋白表达。
结果与结论：①动物实验：苏木精-伊红染色结果显示，单侧前牙反牙合组髁突软骨纤维层崩解、细胞排列无序，增殖层细胞密集，细胞簇集明显，软骨下骨有炎症细胞浸润；番红O-固绿染色结果显示，单侧前牙反牙合组软骨基质红染范围变小、颜色变浅，潮线不连续。单侧前牙反牙合组髁突软骨中白细胞介素1β、基质金属蛋白酶13的mRNA与蛋白表达均高于对照组(P < 0.05)，Ⅱ型胶原、蛋白聚糖、线粒体肌酸激酶2的mRNA与蛋白表达均低于对照组(P < 0.05)。②细胞实验：模型组髁突软骨细胞中基质金属蛋白酶13蛋白表达高于对照组(P < 0.05)，Ⅱ型胶原蛋白表达低于对照组(P < 0.05)。随着白细胞介素1β处理时间的延长，模型组髁突软骨细胞中线粒体肌酸激酶2蛋白表达逐渐降低。③结果表明：大鼠颞下颌关节骨关节炎中线粒体肌酸激酶2的相对表达量与软骨细胞炎症程度呈负相关关系，推测线粒体肌酸激酶2可能是评估颞下颌关节骨关节炎炎症进展状态的重要指标。
https://orcid.org/0009-0008-7326-6973(尼格阿依·艾合麦提)

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

关键词: 颞下颌关节, 骨关节炎, 线粒体肌酸激酶2, 髁突软骨, 白细胞介素1β, 工程化组织构建

Abstract: BACKGROUND: Due to its unknown etiology and the lack of definitive curative treatments, management of temporomandibular joint osteoarthritis primarily focuses on symptom relief. Therefore, the identification of effective early molecular diagnostic biomarkers or potential therapeutic targets holds great significance.
OBJECTIVE: To investigate the expression of mitochondrial creatine kinase 2 in temporomandibular joint osteoarthritis in rats and its role in the progression of inflammation. 
METHODS: (1) Animal experiment: Twenty Sprague-Dawley rats were randomly divided into control and unilateral anterior crossbite groups (n=10). A rat model of temporomandibular joint osteoarthritis was made in the unilateral anterior crossbite group. Four weeks after modeling, histological evaluations, including hematoxylin-eosin and Safranin O-fast green staining, were performed to assess pathological changes in the cartilage and subchondral bone of the mandibular condyle. Quantitative real-time PCR and immunohistochemical staining were utilized to detect the mRNA and protein expression levels of interleukin-1β, matrix metalloproteinase 13, type II collagen, aggrecan, and mitochondrial creatine kinase 2 in condylar cartilage. (2) Cell experiment: Passage 3 condylar cartilage cells from Sprague-Dawley rats were divided into control group and model group. Cells in the control group were routinely cultured, while an inflammation model of condylar cartilage cells was established with interleukin 1β in the model group were treated with interleukin-1β to induce inflammatory cell models. After 24 hours of interleukin-1β treatment, western blot was used to evaluate the expression of matrix metalloproteinase 13, type II collagen proteins in chondrocytes. Western blot was also used to detect the protein expression of mitochondrial creatine kinase 2 in the model group at 0, 12, 24, 48, and 72 hours after treatment with interleukin-1β. 
RESULTS AND CONCLUSION: (1) Animal experiment: The results of hematoxylin-eosin staining showed that the unilateral anterior crossbite group exhibited disintegration of the fibrous layer in the cartilage of the mandibular condyle, disorganization of chondrocyte hierarchy, dense cellularity of the proliferative layer, obvious cell clustering, and infiltration of inflammatory cells in the subchondral bone. The results of Safranin O-fast green staining showed that in the cartilage matrix in the unilateral anterior crossbite group, the red stain became less extensive and lighter in color, and the tidal line was discontinuous. The mRNA and protein expression levels of interleukin-1β and matrix metalloproteinase 13 were elevated in the unilateral anterior crossbite group compared with the control group (P < 0.05). Conversely, the relative expression of type II collagen, aggrecan, and mitochondrial creatine kinase 2 mRNA and protein decreased (P < 0.05). (2) Cell experiment: Compared with the control group, the protein expression of matrix metalloproteinase 13 in the experimental group was significantly increased (P < 0.05), while the protein expression of type II collagen was decreased (P < 0.05). With prolonged interleukin-1β treatment, the protein expression of mitochondrial creatine kinase 2 in condylar chondrocytes of the model group gradually decreased. All the results indicate that the relative expression level of mitochondrial creatine kinase 2 was negatively correlated with the degree of chondrocyte inflammation in the rat model of temporomandibular joint osteoarthritis. Therefore, it is reasonable to infer that mitochondrial creatine kinase 2 is an important indicator for assessing the progression of inflammation in temporomandibular joint osteoarthritis.


Key words: temporomandibular joint, osteoarthritis, mitochondrial creatine kinase 2, condylar cartilage, interleukin-1β, engineered tissue construction

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