MicroRNA-214调控骨关节炎软骨和软骨下骨代谢的机制  

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

作　　者：天生 王玺 王永成 刘亚宁 阳鸿全 Tian Sheng;Wang Xi;Wang Yongcheng;Liu Yaning;Yang Hongquan(Graduate School of Baotou Medical College,Inner Mongolia University of Science and Technology,Baotou 014000,Inner Mongolia Autonomous Region,China;Guang’an District People’s Hospital of Guang’an City,Guang’an 638000,Sichuan Province,China;Inner Mongolia Medical University,Hohhot 010000,Inner Mongolia Autonomous Region,China;Orthopedic Center,Inner Mongolia People’s Hospital,Hohhot 010000,Inner Mongolia Autonomous Region,China)

机构地区：[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

出　　处：《中国组织工程研究》2025年第12期2466-2474,共9页Chinese Journal of Tissue Engineering Research

基　　金：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。

摘　　要：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�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 Experi

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 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R684.3