手术诱导大鼠中重度膝骨关节炎模型的建立与评价  

Establishment and Evaluation of a Moderate-to-Severe Knee Osteoarthritis Model in Rats Induced by Surgery

作　　者：孙效容 苏丹 贵文娟 陈玥 SUN Xiaorong;SU Dan;GUI Wenjuan;CHEN Yue(Chengdu Bio-HT Co.,Ltd.,Chengdu 610041,China;State Key Laboratory of Biotherapy,Sichuan University,Chengdu 610041,China;General Hospital of Ningxia Medical University,Yinchuan 750004,China)

机构地区：[1]成都合拓创展生物科技有限公司,成都610041 [2]四川大学生物治疗全国重点实验室,成都610041 [3]宁夏医科大学总医院,银川750004

出　　处：《实验动物与比较医学》2024年第6期597-604,共8页Laboratory Animal and Comparative Medicine

基　　金：四川省科技计划项目“科技型企业高级管理人才”(2021JDRC0061)。

摘　　要：目的建立大鼠中重度膝骨关节炎模型,为中重度膝关节炎发病机制及其防治方法研究奠定基础。方法选用30只雄性SD大鼠,随机分为3组:假手术组、模型8周组、模型20周组,每组各10只。模型8周组和模型20周组大鼠通过手术切断右膝关节前后交叉韧带和内侧副韧带,去除内外侧半月板,术后大鼠可自由活动。假手术组大鼠仅切开皮肤,暴露关节,不进行手术处理。分别于术后8周和20周进行Micro-CT观察,分析大鼠股骨骨质疏松情况;大鼠安乐死后,对膝关节面进行大体观察,并采用Pelletier评分表对关节面软骨进行评分。膝关节取材,进行HE染色和番红O-固绿染色,观察软骨形态变化,并使用改良Mankin's评分法对关节面组织病理情况进行评分。通过免疫组织化学染色法检测Ⅱ型胶原和基质金属蛋白酶13(matrix metalloproteinase 13,MMP13)表达情况,以此反映膝关节软骨的合成分解代谢情况。结果模型8周组和模型20周组大鼠膝关节的关节面软骨破坏严重,Pelletier评分及改良Mankin's评分均明显高于假手术组(均P<0.01)。模型20周组大鼠的Pelletier评分及改良Mankin's评分均明显高于模型8周组(P<0.01)。模型8周组和模型20周组大鼠经Micro-CT观察可见膝关节面不平整,骨赘形成,出现骨质疏松的表现;且模型20周组大鼠的膝关节周围有较多游离体形成。免疫组织化学染色提示模型组大鼠的膝关节组织中MMP13表达量增多,Ⅱ型胶原表达量减少,提示关节软骨合成分解代谢平衡被破坏。结论通过手术切断大鼠膝关节前后交叉韧带和内侧副韧带并去除内外侧半月板的方法可成功构建中重度膝关节炎大鼠模型,影像学检查发现膝关节骨赘、骨质疏松及游离体,病理学观察发现关节软骨减少甚至消失,而且软骨合成分解代谢平衡破坏。Objective To establish a rat model of moderate-to-severe knee osteoarthritis,laying the foundation for studying the pathogenesis of moderate-to-severe knee osteoarthritis and its prevention and treatment methods.Methods Thirty male SD rats were randomly divided into three groups:a sham surgery group,an 8-week model group,and a 20-week model group,with 10 rats in each group.Rats in the 8-week and 20-week model groups underwent surgery to cut the anterior and posterior cruciate ligaments and medial collateral ligament of the right knee joint,and remove the medial and lateral menisci.After surgery,the rats were allowed to move freely.The rats in the sham surgery group had only skin incisions to expose the joint without any surgical treatment.At 8 and 20 weeks post-surgery,Micro-CT scans were performed to analyze the femoral osteoporosis in the rats.After euthanizing the rats,gross observations of the knee joints were made,and the cartilage of the joint surface was scored using the Pelletier scoring system.The knee joints were collected for hematoxylin and eosin(HE)staining and safranin O-fast green staining to observe changes in cartilage morphology.The modified Mankin's scoring system was used to assess the tissue pathology of the joint surface.Immunohistochemical staining was used to detect the expression of type II collagen and matrix metalloproteinase 13(MMP13),reflecting the anabolic and catabolic metabolism of the knee joint cartilage.Results The knee joint cartilage in the 8-week and 20-week model groups was severely damaged,with Pelletier and modified Mankin's scores significantly higher than those in the sham surgery group(both P<0.01).The Pelletier and modified Mankin's scores in the 20-week model group were significantly higher than those in the 8-week model group(P<0.01).Micro-CT observations revealed irregular joint surfaces,osteophyte formation,and signs of osteoporosis in both the 8-week and 20-week model groups,with the 20-week model group showing more loose bodies around the knee joints.Immunohistoc

关 键 词：动物模型 膝骨关节炎 MICRO-CT 组织病理学 大鼠 

分 类 号：R684.3[医药卫生—骨科学] Q95-33[医药卫生—外科学]