Wear debris inhibition of anti-osteoclastogenic signaling by interleukin-6 and interferon-γ -: Mechanistic insights and implications for periprosthetic osteolysis
Wear debris inhibition of anti-osteoclastogenic signaling by interleukin-6 and interferon-γ -: Mechanistic insights and implications for periprosthetic osteolysis
复制标题
DOI：
10.2106/jbjs.e.00711
复制
发表时间：
2006-04-01
影响因子：
5.3
通讯作者：
Purdue, PE
中科院分区：
文献类型：
--
作者：
Rakshit, DS;Ly, K;Purdue, PE
背景：巨噬细胞对磨损碎片的攻击刺激促炎细胞因子的产生，促进假体周围骨溶解。然而，尚不清楚这种作用是否伴随着假体周围组织中可能参与抗破骨细胞活性的其他细胞因子的重编程。在本研究中，我们检测了磨损碎片颗粒抑制两种细胞因子（白细胞介素-6和干扰素- γ）信号传导的能力。方法：在加入细胞因子白介素-6或干扰素- γ之前，用钛或聚甲基丙烯酸甲酯骨水泥颗粒刺激人破骨细胞前体细胞。白细胞介素-6信号是通过测量STAT3信号转导的激活来确定的，使用免疫印迹法和电泳迁移转移法。干扰素- γ信号是通过使用免疫印迹法和电泳迁移位移法测量STAT1的激活和使用实时逆转录-聚合链反应法测量干扰素- γ诱导基因的表达来确定的。通过在这些试验中加入丝裂原激活的蛋白激酶抑制剂，以及通过免疫印迹评估磨损碎片颗粒对丝裂原激活的蛋白激酶的激活，来评估丝裂原激活的蛋白激酶在细胞因子信号传导中的作用。使用实时逆转录-聚合链反应法评估磨损碎片对细胞因子抑制因子SOCS1和SOCS3（以及促炎介质）表达的调节。结果：钛和聚甲基丙烯酸甲酯颗粒均能有效抑制白细胞介素6诱导的人破骨细胞前体细胞中STAT3的激活。钛和聚甲基丙烯酸甲酯激活的p38丝裂原活化蛋白激酶的抑制作用逆转了这些颗粒对白细胞介素-6信号的抑制作用，而ERK和JNK丝裂原活化蛋白激酶（也被两种类型的磨损碎片激活）的抑制作用没有效果。钛和聚甲基丙烯酸甲酯也都诱导了SOCS3的表达，SOCS3是一种白细胞介素-6信号传导抑制剂。除了对白细胞介素-6信号传导有影响外，钛还能显著抑制干扰素- γ诱导的STAT1激活和干扰素- γ诱导基因的表达，而聚甲基丙烯酸甲酯对干扰素- γ信号传导没有影响。结论：钛能抑制人破骨细胞前体细胞中的干扰素- γ和白细胞介素-6信号，而聚甲基丙烯酸甲酯骨水泥只抑制后者。磨损颗粒对白细胞介素-6的抑制具体涉及p38丝裂原活化蛋白激酶的激活，并伴随着白细胞介素-6信号抑制剂SOCS3的大量诱导。相反，钛对干扰素- γ信号的抑制不依赖于有丝分裂原激活的蛋白激酶的激活，并且只伴随干扰素- γ抑制剂SOCS1的适度诱导。临床意义：磨损碎片在假体周围骨溶解发展中的关键作用可能包括抑制抗炎/抗破骨细胞因子信号，以及诱导促炎介质。因此，增强这些“保护性”信号通路的策略可能具有治疗假体周围骨溶解的治疗潜力。
Background: Wear debris challenge of macrophages provokes the generation of proinflammatory cytokines, which contribute to periprosthetic osteolysis. However, it is not known whether this effect is accompanied by reprogramming of other cytokines present within the periprosthetic tissue that may be involved in anti-osteoclastogenic activities. In the present study, we examined the ability of wear debris particles to inhibit the signaling of two such cytokines, interleukin-6 and interferon-gamma.Methods: Human osteoclast precursor cells were challenged with particles of titanium or polymethylmethacrylate bone cement prior to the addition of the cytokines interleukin-6 or interferon-gamma. Interleukin-6 signaling was determined by measuring the activation of STAT3 signal transduction with use of immunoblotting and electrophoretic mobility shift assays. Interferon-gamma signaling was determined by measuring the activation of STAT1 with use of immunoblotting and electrophoretic mobility shift assays and by measuring the expression of interferon-gamma-inducible genes with use of real-time reverse transcription-polymerise chain reaction assays. Involvement of mitogen-activated protein kinases in cytokine signaling was assessed by including mitogen-activated protein kinase inhibitors in these assays and also by means of immunoblot assessment of mitogen-activated protein kinase activation by wear debris particles. Wear debris modulation of expression of the cytokine suppressors SOCS1 and SOCS3 (as well as pro-inflammatory mediators) was assessed with use of real-time reverse transcription-polymerise chain reaction assays.Results: Both titanium and polymethyl methacrylate particles potently inhibited interleukin-6-induced STAT3 activation in human osteoclast precursor cells. Inhibition of p38 mitogen-activated protein kinase, which is activated by titanium and polymethylmethacrylate, reversed the inhibitory effects of these particles on interleukin-6 signaling, whereas inhibition of ERK and JNK mitogen-activated protein kinases (which are also activated by both types of wear debris) had no effect. Titanium and polymethylmethacrylate also both induced expression of SOCS3, an inhibitor of interleukin-6 signaling. In addition to its effects on interleukin-6 signaling, titanium also profoundly inhibited the interferon-gamma-induced activation of STAT1 and the expression of interferon-gamma-inducible genes, whereas polymethylmethacrylate had no effect on interferon-gamma signaling.Conclusions: Titanium inhibits both interferon-gamma and interleukin-6 signaling in human osteoclast precursor cells, whereas polymethylmethacrylate bone cement inhibits only the latter. Wear particle inhibition of interleukin-6 specifically involves the activation of p38 mitogen-activated protein kinase and is accompanied by substantial induction of SOCS3, an inhibitor of interleukin-6 signaling. In contrast, titanium inhibition of interferon-gamma signaling is not dependent on mitogen-activated protein kinase activation and is accompanied by only modest induction of the interferon-gamma inhibitor SOCS1.Clinical Relevance: The critical role of wear debris in the development of periprosthetic osteolysis likely involves the inhibition of anti-inflammatory/anti-osteoclastogenic cytokine signaling in addition to the well-established induction of pro-inflammatory mediators. Strategies to augment these "protective" signaling pathways may therefore have therapeutic potential for the treatment of periprosthetic osteolysis.