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  

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

作　　者：尼格阿依·艾合麦提 伊丽丹娜·地里夏提 安玮[1,2] 买买提吐逊·吐尔地 Nigeayi·Aihemaiti;Yilidanna·Dilixiati;An Wei;Maimaitituxun·Tuerdi(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)

机构地区：[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

出　　处：《中国组织工程研究》2025年第32期6877-6884,共8页Chinese Journal of Tissue Engineering Research

基　　金：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~~。

摘　　要：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�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 1βin the model group were treated with interleukin-1βto induce inflammatory cell models.After 24 hours of interleukin-1βtreatment,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 ex

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 

分 类 号：R459.9[医药卫生—治疗学] R319[医药卫生—临床医学] R782.6