中国组织工程研究

中国组织工程研究 ›› 2025, Vol. 29 ›› Issue (12): 2466-2474.doi: 10.12307/2025.389

• 软骨组织构建 cartilage tissue construction • 上一篇    下一篇

MicroRNA-214调控骨关节炎软骨和软骨下骨代谢的机制

天  生1，王  玺2，3，王永成4，刘亚宁3，阳鸿全1   

  1. 1内蒙古科技大学包头医学院研究生院，内蒙古自治区包头市  014000；2广安市广安区人民医院，四川省广安市  638000；3内蒙古医科大学，内蒙古自治区呼和浩特市  010000；4内蒙古自治区人民医院骨科中心，内蒙古自治区呼和浩特  010000


  • 收稿日期:2024-03-19 接受日期:2024-06-07 出版日期:2025-04-28 发布日期:2024-09-09
  • 通讯作者: 王永成，博士，主任医师，硕士生导师，内蒙古自治区人民医院骨科中心，内蒙古自治区呼和浩特市 010000
  • 作者简介:天生，男，1998年生，内蒙古自治区通辽市人，蒙古族，包头医学院在读硕士，主要从事关节骨科方面的研究。
  • 基金资助:
    内蒙古自然科学基金项目(2020MS08054)，项目负责人：王永成；内蒙古医科大学联合项目(YKD2021LH037)，项目负责人：王永成；2022年度内蒙古自治区卫生健康科技计划项目(202201013)，项目负责人：王永成

Mechanism underlying microRNA-214 regulation of cartilage and subchondral bone metabolism in osteoarthritis

Tian Sheng1, Wang Xi2, 3, Wang Yongcheng4, Liu Yaning3, Yang Hongquan1   

  1. 1Graduate School of Baotou Medical College, Inner Mongolia University of Science and Technology, Baotou 014000, Inner Mongolia Autonomous Region, China; 2Guang’an District People’s Hospital of Guang’an City, Guang’an 638000, Sichuan Province, China; 3Inner Mongolia Medical University, Hohhot 010000, Inner Mongolia Autonomous Region, China; 4Orthopedic Center, Inner Mongolia People’s Hospital, Hohhot 010000, Inner Mongolia Autonomous Region, China
  • Received:2024-03-19 Accepted:2024-06-07 Online:2025-04-28 Published:2024-09-09
  • Contact: Wang Yongcheng, MD, Chief physician, Master’s supervisor, Orthopedic Center, Inner Mongolia People’s Hospital, Hohhot 010000, Inner Mongolia Autonomous Region, China
  • About author:Tian Sheng, Master candidate, Graduate School of Baotou Medical College, Inner Mongolia University of Science and Technology, Baotou 014000, Inner Mongolia Autonomous Region, China
  • Supported by:
     Inner Mongolia Natural Science Foundation, No. 2020MS08054 (to WYC); Joint Project of Inner Mongolia Medical University, No. YKD2021LH037 (to WYC); 2022 Autonomous Region Health and Wellness Science and Technology Program Project, No. 202201013 (to WYC)

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摘要：


文题释义：
microRNA-214(miR-214)：属于微小RNA 家族的一员，是一种非编码RNA，长度通常约为22个核苷酸，在细胞内发挥着重要的调节作用。它可以通过与特定的信使RNA结合，导致这些 mRNA的降解或抑制其翻译，从而调节基因的表达。具体来说，miR-214可以参与多种生物学过程，包括细胞增殖、分化、凋亡、细胞代谢等。
腺病毒：是一种常见病毒，可感染包括人类在内的多种生物。科学家们利用腺病毒的特性对其进行改造，使腺病毒成为基因转移载体。在基因治疗和疫苗研究中，通常将治疗或疫苗抗原基因插入腺病毒载体，然后将这些基因传递给靶细胞，使其表达相应的蛋白质，从而治疗疾病或诱导免疫反应。

背景：microRNA-214(miR-214)在骨质疏松中的作用国内外已有相关报道，而miR-214与骨关节炎关节软骨及软骨下骨退变之间的相互关系尚不清楚。
目的：探讨miR-214与小鼠膝骨关节炎软骨及软骨下骨退变之间存在的关系。
方法：取30只C57BL/6J小鼠随机分组：①实验一：分为假手术组和内侧半月板失稳组(n=3)，分别进行苏木精-伊红染色和荧光定量PCR检测miR-214基因的表达变化；②实验二：分为假手术组、内侧半月板失稳组、内侧半月板失稳+空载腺病毒组(空载组)、内侧半月板失稳+
miR-214拮抗剂过表达腺病毒组(拮抗剂组)(n=6)，术后4周各组分别取软骨组织，进行苏木精-伊红、番红O固绿、甲苯胺蓝染色分析；荧光定量PCR、Western blot检测关节软骨中相关因子的表达情况。
结果与结论：①实验一中，苏木精-伊红染色结果显示，与假手术组相比，内侧半月板失稳组可见软骨退变；荧光定量PCR检测结果显示，所有样本均有miR-214表达，而内侧半月板失稳组软骨样本中miR-214的表达水平明显高于假手术组(P < 0.05)；②实验二中，苏木精-伊红染色、番红O固绿染色、甲苯胺蓝染色结果显示，拮抗剂组软骨退变程度低于内侧半月板失稳组；腺病毒验证PCR检测结果显示，空载组软骨中miR-214表达水平高于拮抗剂组(P < 0.05)；③实验二中，X射线片显示，内侧半月板失稳组和空载组呈典型骨关节炎影像学变化，拮抗剂组关节退行性病变程度相对较轻；Mirco-CT检测结果显示，拮抗剂组在注入miR-214拮抗剂后，骨小梁结构模型指数变小，各项数据整体好于内侧半月板失稳组及空载组；④实验二Western blot检测结果显示，内侧半月板失稳组、空载组软骨标本中软骨相关因子Ⅱ型胶原α1、性别决定区Y框转录因子9、Runt相关转录因子2、骨桥蛋白的相对表达水平低于假手术组及拮抗剂组(P < 0.05)；而金属基质蛋白酶13的相对表达水平在内侧半月板失稳组、空载组高于假手术组及拮抗剂组(P < 0.05)；⑤实验二荧光定量PCR检测结果显示，与假手术组比较，肿瘤坏死因子α和白细胞介素6 mRNA的相对表达量在内侧半月板失稳组、空载组中表达相对较高，拮抗剂组中表达相对较低
(P < 0.05)；内侧半月板失稳组、空载组高于拮抗剂组(P < 0.05)；⑥结果表明，骨关节炎小鼠模型的关节软骨中miR-214表达水平明显升高，提示miR-214表达水平的升高与骨关节炎有密切联系；而在骨关节炎小鼠模型膝关节腔内注入miR-214拮抗剂，可以延缓关节软骨退化、促进软骨下骨重塑，改善骨关节炎进展。
https://orcid.org/0009-0006-1407-3173(天生)

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: microRNA-214, 骨关节炎, 关节软骨, 肿瘤坏死因子α, 白细胞介素6, 软骨下骨

Abstract: BACKGROUND: The role of microRNA-214 in osteoporosis has been reported both at home and abroad, whereas the interrelationship between microRNA-214 and osteoarthritic articular cartilage and subchondral bone degeneration is unclear. 
OBJECTIVE: To investigate the relationship between microRNA-214 and cartilage and subchondral bone degeneration in mice with knee osteoarthritis. 
METHODS: Thirty C57BL/6J mice were randomly grouped: Experiment 1: sham operation group and medial meniscus destabilization group (n=3 per group) underwent hematoxylin-eosin staining and qPCR to detect changes in microRNA-214 gene expression; Experiment 2: sham operation group, medial meniscus destabilization group, medial meniscus destabilization+null-loaded adenovirus group (null-loaded group), and medial meniscus destabilization+microRNA-214 antagonist overexpression adenovirus group (antagonist group; n=6 per group). Cartilage tissues were taken from each group 4 weeks after surgery, and stained with hematoxylin-eosin, safranin O-fast green, and toluidine blue. qPCR and western blot were used to detect the expression of related factors in articular cartilage.
RESULTS AND CONCLUSION: (1) In Experiment 1, hematoxylin-eosin staining results showed that cartilage degeneration was visible in the medial meniscus destabilization group compared with the sham operation group. qPCR assay results showed that microRNA-214 was expressed in all the samples, and the expression level of microRNA-214 in cartilage samples of the medial meniscus destabilization group was significantly higher than that of the sham operation group (P < 0.05). (2) In Experiment 2, the results of hematoxylin-eosin staining, safranin O-fast green staining, and toluidine blue staining showed that the degree of cartilage degeneration in the antagonist group was significantly reduced compared with the medial meniscus destabilization group. Adenovirus-validated PCR assay showed that the microRNA-214 expression level in cartilage tissue was higher in the null-loaded group than in the antagonist group (P < 0.05). (3) In Experiment 2, X-ray results showed typical osteoarthritis imaging changes in the medial meniscus destabilization group and null-loaded group, while the degree of degenerative joint lesions was relatively mild in the antagonist group. The results of microcomputed tomography showed that after injection of microRNA-214 antagonist, trabecular structure model index became smaller in the antagonist group, and the data were better than those of the medial meniscus destabilization group and null-loaded group. (4) In Experiment 2, western blot results showed that The relative expression levels of cartilage-associated factor type II collagen α1, sex-determining region Y-box 9, Runt-associated transcription factor 2, and osteopontin in cartilage specimens of the medial meniscus destabilization group and the null-loaded group were lower than that in the sham operation group and the antagonist group (P < 0.05), whereas the relative expression level of matrix metalloproteinase 13 was higher in the medial meniscus destabilization group and the null-loaded group than the sham operation group and the antagonist group (P < 0.05). (5) In Experiment 2, PCR results indicated that the relative mRNA expression of tumor necrosis factor-α and interleukin-6 was relatively higher in the medial meniscus destabilization group and null-loaded group, but relatively lower in the antagonist group, as compared with the sham operation group (P < 0.05). The relative mRNA expression of tumor necrosis factor-α and interleukin-6 was also higher in the medial meniscus destabilization group and the null-loaded group compared with the antagonist group (P < 0.05). To conclude, the expression level of microRNA-214 in articular cartilage was elevated in the mouse osteoarthritis model, suggesting that the elevated expression level of microRNA-214 is closely related to osteoarthritis; and injection of microRNA-214 antagonist into the knee joint cavity of the mouse osteoarthritis model could delay articular cartilage degradation, promote subchondral bone remodeling, and ameliorate the progression of osteoarthritis.

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

Key words: microRNA-214, osteoarthritis, articular cartilage, tumor necrosis factor-α, interleukin-6, subchondral bone

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