神经生长因子抗体在小鼠膝关节炎疼痛模型中的作用研究  

The Effect of Nerve Growth Factor Antibody in Mouse Knee Arthritis Pain Model

作　　者：陈永锋 冯亚非 宋和强 王鹏 燕明 CHEN Yong-feng;FENG Ya-fei;SONG He-qiang;WANG Peng;YAN Ming(Department of Orthopedics,Xijing Hospital,Air Force Military Medical University,Xi'an,Shaanxi,710032,China)

机构地区：[1]空军军医大学西京医院骨科,陕西西安710032

出　　处：《现代生物医学进展》2021年第21期4038-4044,共7页Progress in Modern Biomedicine

基　　金：国家自然科学基金面上项目(81871799)。

摘　　要：目的:探究神经生长因子(Nerve growth factor,NGF)抗体L148M在碘乙酸(Monoiodoacetate,MIA)诱导的膝关节炎(Kee osteoarthritis,KOA)小鼠模型中的作用机制。方法:随机将8周龄C57BL/6雄性小鼠分为对照组、MIA组和L148M组。采用关节腔注射20 mg/m L MIA诱导KOA小鼠模型。手术后2周,L148M组小鼠给予腹腔注射L148M(10 mg/kg)处理。通过苏木精-伊红(HE)染色、番红O染色、OARSI评分和Micro-CT分析评估小鼠膝关节软骨及软骨下骨组织形态学变化。通过qRT-PCR检测CCR2、MCP1、MMP-1、MMP-3、MMP-13、COL10、IL-1β和TNF-α的m RNA表达水平。通过Western blotting检测INOS、COX-2、Collagen II和Aggrecan的蛋白表达水平。通过免疫组织化学法分析VEGFA和Ang-1的蛋白表达水平。通过Micro-CT血管造影分析软骨下骨血管形成数量。结果:HE染色结果显示,L148M组小苏软骨细胞数和关节软骨厚度均高于MIA组。番红O染色显示,L148M组小鼠基质降解小于MIA组。L148M组OARSI评分显著低于MIA组(P<0.05)。micro-CT扫描结果表明,L148M组小鼠软骨和软骨下骨结构完整,没有明显病理损伤。qRT-PCR检测结果显示,与MIA组相比,L148M组小鼠COL10、MMP1、MMP3和MMP-13表达水平均显著降低(P<0.05)。Western blotting检测结果表明,与MIA组相比,L148M组小鼠Collagen II和Aggrecan表达水平升高,INOS和COX-2表达水平降低(P<0.05)。疼痛行为学分析显示,与MIA组相比,L148M组小鼠行进距离和机械刺激反应阈值均降低(P<0.05)。micro-CT血管造影分析显示,L148M小鼠组微血管数量和体积明显低于MIA组(P<0.05)。免疫组化检测显示,L148M组小鼠VEGFA和Ang-1蛋白表达水平均显著低于MIA组(P<0.05)。结论:L148M通过抑制软骨下骨异常血管生成,缓解关节炎症疼痛;并通过抑制炎症细胞因子表达,减轻软骨和软骨下骨病理损伤。Objective:To explore the molecular mechanism of the protective effect of Nerve growth factor(NGF)antibody L148 M in a mouse model of Kio osteoarthritis(KOA)induced by iodoacetic acid(MIA).Methods:Eight-week-old male C57 BL/6 male mice were randomly divided into a control group,MIA group,and L148 M group.KOA mouse model was induced by injecting 20 mg/m L MIA into joint cavity.Two weeks after the operation,mice in the L148 M group were intraperitoneally injected with L148 M(10 mg/kg).Hematoxylin-eosin(HE)staining,safranin O staining,OARSI score,and Micro-CT analysis were used to observe the morphological changes of knee joint cartilage and subchondral bone in mice.The expression levels of CCR2,MCP1,MMP-1,MMP-3,MMP-13,COL10,IL-1βand TNF-αm RNA were detected by qRT-PCR.The expression levels of INOS,COX-2,Collagen II and Aggrecan were detected by Western blotting.The expression of VEGFA and Ang-1 protein was analyzed by immunohistochemistry.Micro-CT angiography analyzes the number of new blood vessels in subchondral bone.Results:HE staining results showed that the number of chondrocytes and cartilage thickness in the L148 M group were higher than those in the MIA group.Safranin O staining showed that the matrix degradation of the L148 M group was less than that of the MIA group.The OARSI score of the L148 M group was significantly lower than that of the MIA group(P<0.05).The results of micro-CT scan showed that the cartilage and subchondral bone of the L148 M group were intact and there was no obvious pathological damage.The results of qRT-PCR showed that compared with the MIA group,the m RNA expression level of COL10,MMP1,MMP3 and MMP-13 in the L148 M group were significantly reduced(P<0.05).Western blotting results showed that compared with the MIA group,the protein expression levels of Collagen II and Aggrecan were increased in the L148 M group,and the protein expressions level of INOS and COX-2 were decreased(P<0.05).Pain behavior analysis showed that compared with the MIA group,the travel distance and mechanica

关 键 词：膝关节炎 神经生长因子抗体 软骨基质 血管形成 

分 类 号：R-33[医药卫生] R684.3