The pro-inflammatory role of high-mobility group box 1 protein (HMGB-1) in photoreceptors and retinal explants exposed to elevated pressure
The pro-inflammatory role of high-mobility group box 1 protein (HMGB-1) in photoreceptors and retinal explants exposed to elevated pressure
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DOI：
10.1038/labinvest.2015.156
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发表时间：
2016-04-01
影响因子：
5
通讯作者：
Thanos, Solon
中科院分区：
文献类型：
--
作者：
Boehm, Michael R. R.;Schallenberg, Maurice;Thanos, Solon
确定高迁移率族蛋白1（HMGB-1）在高压诱导的神经变性、再生和炎症的细胞和组织模型中的作用。将小鼠视网膜光感受器衍生细胞（661 W）和视网膜外植体在高压下或在重组HMGB-1（rHMGB-1）存在下孵育，以研究光感受器的反应机制。采用免疫组化、western blotting和实时荧光定量PCR检测免疫因子的表达水平（例如，HMGB-1，晚期糖基化终产物受体（receptor for advanced glycation end products，HMG）），Toll样受体2和4（TLR-2，TLR-4），骨化相关因子（例如，B细胞淋巴瘤2（Bcl-2）、Bcl-2相关死亡启动子（Bad））以及细胞因子表达（例如，肿瘤坏死因子α（TNF-α）、白细胞介素（IL）-4、IL-6和血管内皮生长因子（VEGF））。数据显示，在暴露于高压的661 W细胞中，HMGB-1及其受体TLR-2和TLR-4、TNF-α以及促凋亡因子（例如，Bad）的表达增加，以及凋亡。rHMGB-1与661 W细胞共培养后，促凋亡蛋白Bad和Caspase-3的表达水平增加，导致细胞凋亡。细胞因子阵列研究显示，在用rHMGB-1孵育661 W细胞后，TNF-α、IL-4、IL-6和VEGF的释放增加。在暴露于rHMGB-1或升高的压力的视网膜外植体中发现HMGB-1、TLR-2和Bcl-2以及抗凋亡Bcl-2表达水平的上调。结果表明，HMGB-1促进炎症反应，并介导光感受器和视网膜稳态的病理凋亡。HMGB-1可能在眼内压升高条件下视网膜细胞的持续损伤中起关键作用。
To determine the role of high-mobility group box 1 protein (HMGB-1) in cellular and tissue models of elevated pressure induced neurodegeneration, regeneration, and inflammation. Mouse retinal photoreceptor-derived cells (661W) and retinal explants were incubated either under elevated pressure or in the presence of recombinant HMGB-1 (rHMGB-1) to investigate the mechanisms of response of photoreceptors. Immunohistochemistry, western blotting, and the quantitative real-time PCR were used to examine the expression levels of immunological factors (eg, HMGB-1, receptor for advanced glycation end products (RAGE)), Toll-like receptors 2 and 4 (TLR-2, TLR-4), apoptosis-related factors (eg, B-cell lymphoma 2 (Bcl-2), Bcl-2-associated death promoter (Bad)) as well as cytokine expression (eg, tumor necrosis factor alpha (TNF-alpha), interleukin (IL)-4, IL-6, and vascular endothelial growth factor (VEGF)). The data revealed increased the expression of HMGB-1 and its receptors RAGE, TLR-2, and TLR-4, and TNF-a as well as pro-apoptotic factors (eg, Bad) as well as apoptosis in 661W cells exposed to elevated pressure. Co-cultivation of 661W cells with rHMGB-1 increased the expression levels of pro-apoptotic Bad and cleaved Caspase-3 resulting in apoptosis. Cytokine array studies revealed an increased release of TNF-a, IL-4, IL-6, and VEGF after incubation of 661W cells with rHMGB-1. Upregulation of HMGB-1, TLR-2, and RAGE as well as anti-apoptotic Bcl-2 expression levels was found in the retinal explants exposed to rHMGB-1 or elevated pressure. The results suggest that HMGB-1 promotes an inflammatory response and mediates apoptosis in the pathology of photoreceptors and retinal homeostasis. HMGB-1 may have a key role in ongoing damage of retinal cells under conditions of elevated intraocular pressure.