Leech Michelle, Lacey Derek, Xue Jin Rong, Santos Leilani, Hutchinson Paul, Wolvetang Ernst, David John R, Bucala Richard, Morand Eric F
Centre for Inflammatory Diseases, Monash University Department of Medicine, Monash Medical Centre, Locked Bag Number 29, Clayton, Melbourne, Victoria 3168, Australia.
Arthritis Rheum. 2003 Jul;48(7):1881-9. doi: 10.1002/art.11165.
To study the capacity of macrophage migration inhibitory factor (MIF) to regulate proliferation, apoptosis, and p53 in an animal model of rheumatoid arthritis (RA) and in fibroblast-like synoviocytes (FLS) from humans with RA.
Antigen-induced arthritis (AIA) was induced in MIF(-/-) mice and littermate controls. FLS were obtained from patients with RA. Western blotting and immunohistochemistry were used to measure p53 in cells and tissues. Apoptosis was detected in cells by flow cytometry using TUNEL and annexin V/propidium iodide labeling. Apoptosis in tissue was detected using TUNEL. Proliferation was assessed in cultured cells and tissue by (3)H-thymidine incorporation and Ki-67 immunostaining, respectively.
MIF inhibited p53 expression in human RA FLS. Levels of p53 were correspondingly increased in MIF(-/-) mouse tissues and cells. Spontaneous and sodium nitroprusside-induced apoptosis were significantly increased in MIF(-/-) cells. In vitro exposure of FLS to MIF reduced apoptosis and significantly induced FLS proliferation. Synoviocyte proliferation in MIF(-/-) mice was correspondingly reduced. A decrease in the severity of AIA in MIF(-/-) mice was associated with an increase in p53 and apoptosis in synovium. Evidence of in situ proliferation was scant in this model, and no difference in in situ proliferation was detectable in MIF(-/-) mice compared with wild-type mice.
These results indicate a role for MIF in the regulation of p53 expression and p53-mediated events in the inflamed synovium and support the hypothesis that MIF is of critical importance in the pathogenesis of RA.
在类风湿关节炎（RA）动物模型以及来自RA患者的成纤维样滑膜细胞（FLS）中，研究巨噬细胞移动抑制因子（MIF）调节细胞增殖、凋亡及p53的能力。
在MIF基因敲除（MIF(-/-)）小鼠及其同窝对照小鼠中诱导抗原诱导性关节炎（AIA）。从RA患者获取FLS。采用蛋白质免疫印迹法和免疫组织化学法检测细胞和组织中的p53。通过TUNEL法以及膜联蛋白V/碘化丙啶标记，利用流式细胞术检测细胞凋亡。使用TUNEL法检测组织中的凋亡情况。分别通过³H-胸腺嘧啶核苷掺入法以及Ki-67免疫染色评估培养细胞和组织中的增殖情况。
MIF抑制人RA FLS中p53的表达。在MIF(-/-)小鼠的组织和细胞中，p53水平相应升高。MIF(-/-)细胞中自发性及硝普钠诱导的凋亡显著增加。FLS在体外暴露于MIF可减少凋亡并显著诱导FLS增殖。MIF(-/-)小鼠中的滑膜细胞增殖相应减少。MIF(-/-)小鼠中AIA严重程度的降低与滑膜中p53及凋亡的增加相关。在该模型中，原位增殖的证据很少，与野生型小鼠相比，在MIF(-/-)小鼠中未检测到原位增殖的差异。
这些结果表明MIF在炎症滑膜中p53表达及p53介导的事件调节中发挥作用，并支持MIF在RA发病机制中至关重要这一假说。
