摘要
目的探讨铜离子螯合剂四硫代钼酸铵(TTM)缓解类风湿关节炎(RA)小鼠的效果。方法24只雄性浅棕色非Agouti基因/1小鼠分为4组(n=6):空白对照组(Ctrl组)、胶原诱导的关节炎(CIA)+磷酸盐缓冲液(PBS)处理组(PBS组)、CIA+TTM处理组(TTM组)、CIA+铜离子载体Elesclomol(Eles)处理组(Eles组)。处理后1周观察小鼠足趾肿胀情况,每周评价1次小鼠关节炎临床积分,小鼠取材前对足趾机械痛进行检测,Catwalk步态分析系统测定小鼠步态,苏木素-伊红(HE)染色评价小鼠足趾病理形态变化,免疫组织化学法检测小鼠足趾中基质金属蛋白酶-3(MMP-3),免疫荧光检测CD31和血管内皮生长因子(VEGF)表达,Luminex技术检测血清炎性因子改变。结果与Ctrl组相比,PBS组和Eles组小鼠足趾关节红肿或畸形,关节炎临床积分明显升高,关节组织骨破坏、滑膜增生及炎性细胞浸润,病理改变明显,小鼠的MMP-3表达显著增加,血清中单核细胞趋化蛋白-1(MCP-1)、白细胞介素(IL)-1β、IL-9以及干扰素-γ(IFN-γ)等炎症因子表达水平明显上升,而抗炎因子IL-10表达水平降低,同时CD31及VEGF因子表达均显著增强。与PBS组比较,TTM组小鼠足趾关节肿胀变形程度显著改善,关节炎临床积分降低,小鼠关节骨破坏程度、炎细胞浸润及滑膜增生,血清MCP-1、IL-1β、IL-9、IFN-γ表达水平明显降低,IL-10水平有上升趋势;与Ctrl组相比,TTM组小鼠MMP-3、CD31、VEGF因子表达无显著差异。结论TTM能够通过同时靶向多种炎症因子、VEGF和骨破坏介质来多重阻断滑膜炎症、血管生成和骨破坏,从而缓解CIA诱导的RA小鼠关节组织病理损伤。
Objective:To investigate the therapeutic effects of the copper chelator ammonium tetrathiomolybdate(TTM)on rheumatoid arthritis(RA)in a mouse model.Methods:Twenty-four male dilution brown non-Agoutia/1 mice were randomly divided into 4 groups:a blank control group(Ctrl group,n=6),a collagen-induced arthritis(CIA)+phosphate buffer saline(PBS)treatment group(PBS group,n=6),a CIA+TTM treatment group(TTM group,n=6),and a CIA+Elesclomol treatment group(Eles group,n=6).Eles,a copper ion carrier,served as a control for administration of TTM,a copper ion chelator.One week after treatment,the swelling of mouse paw was observed,and the clinical scoring of the arthritis in mice was evaluated once a week.Paw mechanical pain detection was performed and photographs were taken to observe the severity of paw swelling before the mice were sacrificed.Catwalk gait analysis system was used to evaluate the gait changes in mice.HE and saffron O solid green staining were used to evaluate pathomorphologic changes in the mice knee joints and paws.Immunostaining techniques were used to detect the protein expression of MMP3,CD31,and VEGF in the mice paws.Luminex technology was used to detect alterations in the serum inflammatory factors.Results:Compared with the Ctrl group,in the PBS and Eles groups,the joints were red,swollen and deformed;the arthritis clinical scores were significantly higher;the bone destruction,synovial hyperplasia and inflammatory cell infiltration and pathological changes in the joint tissues were obvious;the expression levels of inflammatory factors,such as serum MCP-1,IL-1β,IL-9,and IFN-γ,were significantly higher while the expression level of IL-10 was significantly lower.Simultaneously,the expression of CD31 and VEGF factors was significantly enhanced.Compared with the PBS group,in the TTM group,the joint swelling and deformation were significantly improved,the arthritis clinical score was reduced,and the joint bone destruction,inflammatory cell infiltration and synovial hyperplasia were alleviated,and the levels of serum MCP-1,IL-1β,IL-9 and IFN-γwere significantly decreased while the level of anti-inflammatory factor IL-10 was increased.There was no significant difference in the expression of MMP-3,CD31 or VEGF factors between the CTRL group and the TTM group.Conclusion:TTM can block synovial inflammation,angiogenesis,and bone destruction multiple times by simultaneously targeting multiple inflammatory factors,VEGF factors,and bone destruction mediators,thereby alleviating the pathological damage to the joint tissues induced by CIA in RA mice.
作者
李哲
邓辉丽
周绪昌
陈曦儿
林彰裕
骆晓飞
魏瑄
倪国新
Li Zhe;Deng Huili;Zhou Xuchang;Chen Xier;Lin Zhangyu;Luo Xiaofei;Wei Xuan;Ni Guoxin(Department of Joint Diseases,Zhengzhou Orthopedic Hospital,Zhengzhou 450052,China;Department of Rehabilitation Medicine,The First Hospital Affiliated to School of Medicine,Xiamen University,Xiamen 361003,China;School of Sports Medicine and Rehabilitation,Beijing Sports University,Beijing 100084,China;College of Sports Medicine,Wuhan Sports University,Wuhan 430079,China)
出处
《中华创伤骨科杂志》
北大核心
2025年第7期620-628,共9页
Chinese Journal of Orthopaedic Trauma
基金
河南省医学科技攻关计划项目(LHGJ20200756)
厦门市留学人才项目(120221361)。
关键词
关节炎
滑膜炎
软骨
四硫代钼酸铵
骨侵蚀
血管侵袭
Arthritis
Synovitis
Cartilage
Ammonium tetrathiomolybdate
Bone erosion
Vascularinvasion
