Involvement of lymphocytes with a Th1 cytokine profile in bone cell damage associated with MMP-9 production in collagen-induced arthritis
Involvement of lymphocytes with a Th1 cytokine profile in bone cell damage associated with MMP-9 production in collagen-induced arthritis
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DOI：
10.1007/s00011-004-1311-z
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发表时间：
2004-12
影响因子：
6.7
通讯作者：
J. Chen;X. Zhang;Q. Xu
中科院分区：
文献类型：
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作者：
J. Chen;X. Zhang;Q. Xu
目的：探讨淋巴细胞是否能直接引起胶原诱导性关节炎（CIA）小鼠骨细胞的破坏及其机制。方法：用鸡II型胶原诱导昆明小鼠关节炎。采用生物学方法测定TNF-α和IFN-γ的活性。采用逆转录聚合酶链反应（RT-PCR）检测TNF-α、IFN-γ、T-bet、Th1特异性转录因子和Fas的mRNA。通过明胶酶谱分析测定基质金属蛋白酶 (MMP)-9。采用3-(4,5-二甲基-2-噻唑基)-2,5-二苯基-2H-四唑溴化物(MTT)法检测脾细胞对骨细胞群的细胞毒性和脾细胞对鸡II型胶原(CII)的增殖作用。结果:鸡CII成功诱导昆明小鼠关节炎,小鼠足爪肿胀明显,具有典型的关节炎特征。与初始小鼠相比，关节炎小鼠血清中IFN-γ和TNF-α的产生增加，并且关节炎小鼠的脾细胞显示出Th1细胞因子谱并在体外直接损伤骨细胞。有趣的是，与 CIA 小鼠的脾细胞一样，经佛波醇 12-肉豆蔻酸酯 13-乙酸酯 (PMA) 处理的幼稚脾细胞也会损伤骨细胞。环孢菌素 A 阻断 CIA 小鼠脾细胞对骨细胞的细胞毒性。只有抗CD4和补体系统共同抑制CIA小鼠脾细胞对骨细胞的作用。只有患有关节炎的小鼠的脾细胞才能针对小鸡 CII 进行增殖。此外，关节炎小鼠脾细胞的培养上清液中 MMP-9 的活性和 NO 的产生量均高于初始小鼠。 PMA 提高了幼稚脾细胞中 MMP-9 的活性，但不提高 NO 的产生。此外，关节炎小鼠关节组织中 TNF-α、IFN-γ 和 MMP-9 的活性高于未处理小鼠的关节组织，但 NO 的产生却较高。体外实验中，TNF-α 和 IFN-γ 显着提高了关节炎小鼠和幼鼠脾细胞中 MMP-9 的活性。此外，只有抗MMP-9而不是抗MMP-2抗体抑制CIA小鼠脾细胞对骨细胞的细胞毒性。结论：TNF-α和IFN-γ等促炎因子升高可能促进局部炎症细胞中MMP-9的活性，而这些炎症细胞实际上参与了CII诱导的关节炎的骨细胞损伤。
Objective:To investigate whether lymphocytes could directly cause the destruction of bone cells in collageninduced arthritis (CIA) mice and the mechanism of this destruction.Methods:Arthritis was induced with chick type II collagen in Kunming mice. The activities of TNF-α and IFN-γ were measured by biological methods. The mRNA of TNF-α, IFN-γ, T-bet, a Th1 specific transcription factor and Fas was detected by reverse transcript-polymerase chain reaction (RT-PCR). Matrix metalloproteinase (MMP)-9 was measured by gelatin zymography assay. The cytotoxicity of spleen cells to bone cell populations and the proliferation of spleen cells against chick type II collagen (CII) were examined by 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2H-tetrazolium bromide (MTT) method.Results:Arthritis was successfully induced by chick CII in Kunming mice, which had remarkable paw swelling and typical features of arthritis. Compared with naive mice, the arthritic mice showed an increased IFN-γ and TNF-α production in serum, and the spleen cells from the arthritic mice displayed a Th1 cytokine profile and directly damaged bone cells in vitro. Interestingly, naive spleen cells treated by phorbol 12-myristate 13-acetate (PMA) also damaged bone cells as did spleen cells from CIA mice. Cyclosporine A blocked the cytotoxicity of spleen cells from CIA mice to bone cells. Only anti-CD4 and complement system together inhibited the effect of spleen cells from CIA mice on bone cells. Only the spleen cells from arthritic mice could proliferate against chick CII. In addition, the culture supernatants from spleen cells of arthritic mice contained a higher activity of MMP-9 and production of NO than those of naive mice. PMA elevated the activity of MMP-9 but not the production of NO in naive spleen cells. Moreover, the activities of TNF-α, IFN-γ and MMP-9 but not the production of NO were higher in joint tissues of arthritic mice than those of naive mice. TNF-α and IFN-γ significantly elevated the activity of MMP-9 in spleen cells from both arthritic and naive mice in vitro. Moreover, only anti-MMP-9 but not anti-MMP-2 antibody inhibited the cytotoxicity of spleen cells from CIA mice to bone cells.Conclusions:The elevated proinflammatory factors such as TNF-α and IFN-γ may promote the MMP-9 activity in local inflammatory cells that are actually involved in the bone cell damage in CII-induced arthritis.