IL-1 and TNF induction of matrix metalloproteinase-3 by c-Jun N-terminal kinase in trabecular meshwork
IL-1 and TNF induction of matrix metalloproteinase-3 by c-Jun N-terminal kinase in trabecular meshwork
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DOI：
10.1167/iovs.05-0451
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发表时间：
2006-04-01
影响因子：
4.4
通讯作者：
Acott, TS
中科院分区：
文献类型：
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作者：
Hosseini, M;Rose, AY;Acott, TS
目的。细胞因子肿瘤坏死因子和白介素1介导基质金属蛋白酶-3的增加发生在小梁网(TM)激光小梁成形术治疗的反应。这种基质金属蛋白酶-3的增加似乎在这种治疗开角型青光眼的疗效中起着关键作用。蛋白激酶C和ERK丝裂原活化蛋白(MAP)激酶是通过这些细胞因子处理TM细胞而产生的基质金属蛋白酶-3(MMP3)信号转导过程中必不可少的信号成分。在这里，我们评估了JNK-MAP激酶通路在这一过程中的作用。方法：用肿瘤坏死因子α、白介素1α或白介素1β处理猪TM细胞。免疫印迹法检测培养上清液中基质金属蛋白酶-3和基质金属蛋白酶-9蛋白水平的变化。评价JNK抑制剂2的作用。检测肿瘤坏死因子α和白介素1α处理后不同时间JNK、c-jun、ATF-2、MKK4和MKK7的磷酸化水平的变化。将2.3kb的基质金属蛋白酶-3启动子片段克隆到分泌型碱性磷酸酶报告载体中。将构建的报告基因与含有JNK显性负性突变体的哺乳动物表达载体共转染TM细胞。结果：肿瘤坏死因子α、IL-1α和IL-10可增加基质金属蛋白酶-3和基质金属蛋白酶-9的蛋白水平，而JNK抑制物2可阻断这些增加。JNK1/2、MKK4、c-jun和ATF-2的磷酸化水平在肿瘤坏死因子α和白介素1α治疗后升高。JNK抑制剂2可阻断这些c-jun和ATF-2的磷酸化增加。显性负性JNK显著降低由这些细胞因子诱导的基质金属蛋白酶-3启动子驱动的报告活性。结论：JNK活性是肿瘤坏死因子α、IL-1α或IL-1β诱导TM细胞产生基质金属蛋白酶-3和基质金属蛋白酶-9所必需的。JNK信号通路以及转录因子c-jun和ATF-2的磷酸化支持该通路在TM对这些细胞因子的反应中诱导MMP3和MMP9的作用。因此，在TM细胞的这一信号事件中，至少有三条独立的信号转导途径是必要的。
PURPOSE. The cytokines TNF and IL-1 mediate the MMP-3 increase that occurs in response to trabecular meshwork (TM) treatment by laser trabeculoplasty. This MMP-3 increase appears to play a key role in the efficacy of this treatment for open-angle glaucoma. Protein kinase C mu and the Erk mitogen-activated protein (MAP) kinases are essential signaling components in transducing MMP-3 increases produced by treatment of TM cells with these cytokines. Here, the involvement of the JNK-MAP kinase pathway in this process was evaluated.METHODS. Porcine TM cells were treated with TNF alpha, IL-1 alpha, or IL-1 beta. Changes in MMP-3 and MMP-9 protein levels in the media were then determined by Western immunoblot. The effect of JNK inhibitor 2 was evaluated. Changes in the level of phosphorylation of JNK, c-Jun, ATF-2, MKK4, and MKK7 were also determined at various times after TNF alpha or IL-1 alpha treatment. A 2.3-kb MMP-3 promoter fragment was cloned into a secreted alkaline phosphatase reporter vector. This reporter construct was cotransfected into TM cells with a mammalian expression vector containing a dominant-negative mutant of JNK. The involvement of JNK activity in the TNF alpha and IL-1 alpha induction of MMP-3 expression was then evaluated.RESULTS. TNF alpha, IL-1 alpha, and IL-10 increase media MMP-3 and MMP-9 protein levels, and JNK inhibitor 2 blocks these increases. JNK1/2, MKK4, c-Jun, and ATF-2 phosphorylation levels increase in response to TNF alpha and IL-1 alpha treatment. JNK inhibitor 2 pretreatment blocks these c-Jun and ATF-2 phosphorylation increases. Dominant-negative JNK dramatically reduces the MMP-3 promoter-driven reporter activity induced by these cytokines.CONCLUSIONS. JNK activity is necessary for the induction of MMP-3 and MMP-9 by TNF alpha, IL-1 alpha, or IL-1 beta in TM cells. Phosphorylation of components of the JNK signaling pathway and of the transcription factors c-Jun and ATF-2 support a role for this pathway in the induction of MMP-3 and MMP-9 in the TM in response to these cytokines. Thus, at least three separate signal transduction pathways are necessary in this signaling event in TM cells.