HMGB1 promotes the activation of NLRP3 and caspase-8 inflammasomes via NF-κB pathway in acute glaucoma.
HMGB1 promotes the activation of NLRP3 and caspase-8 inflammasomes via NF-κB pathway in acute glaucoma.
复制标题
HMGB1通过NF-κB通路促进急性青光眼中NLRP3和caspase-8炎症小体的激活
DOI：
10.1186/s12974-015-0360-2
复制
发表时间：
2015-07-30
影响因子：
9.3
通讯作者：
Zhuo Y
中科院分区：
文献类型：
--
作者：
Chi W;Chen H;Li F;Zhu Y;Yin W;Zhuo Y
急性青光眼是世界上一种严重威胁视力、导致不可逆转失明的原因，其特征是眼内压（IOP）突然大幅升高，随后视网膜神经节细胞（RGC）死亡。本研究旨在探讨高迁移率族蛋白1（HMGB 1）在急性青光眼小鼠模型中的作用。C57 BL/6小鼠经前房穿刺注入平衡盐溶液，使眼压迅速升高至70 mmHg，持续60 min，建立急性青光眼模型。分别于高眼压治疗后6、24、48、72 h及治疗后48 h观察视网膜组织缺血性损伤和RGCs的丢失情况，并观察高眼压治疗后rHMGB 1、HMGB 1抑制剂烟酸（GA）对视网膜组织缺血性损伤的影响。玻璃体内注射NF-κB（NF-κB）抑制剂JSH-23和caspase-8抑制剂Z-IETD-fetamine。采用逆转录和半定量逆转录聚合酶链反应（RT-PCR）、蛋白质印迹法和免疫沉淀法评估核苷酸结合结构域、富含亮氨酸重复序列的蛋白3（NLRP 3）、磷酸化NF-κB p65、caspase-8、caspase-1、凋亡相关斑点样蛋白（ASC）和白细胞介素-1 β（IL-1β）的表达水平。HMGB 1在急性青光眼缺血视网膜组织中的表达早在眼压快速升高后6 h即开始增加。外源性HMGB 1可加重视网膜缺血性损伤、RGC丢失，抑制内源性HMGB 1可显著降低疾病的严重程度。HMGB 1可显著诱导经典NLRP 3、ASC、caspase-1和非经典caspase-8-ASC炎性小体的表达，并促进IL-1β的加工。HMGB 1对NLRP 3炎性体活化和IL-1β产生的影响依赖于NF-κB通路。因此，HMGB 1/caspase-8途径通过NF-κB途径促进IL-1β的加工。这项研究的结果确定了一种新的信号通路，其中HMGB 1响应于急性升高的眼内压，在急性青光眼发展期间激活经典NLRP 3和非经典caspase-8炎性小体并产生IL-1β。这些结果提供了新的见解的先天性反应，有助于急性青光眼的发病机制的理解。
Acute glaucoma is a significantly sight-threatening cause of irreversible blindness in the world characterized by a sudden and substantial intraocular pressure (IOP) increase and subsequent retinal ganglion cell (RGC) death. This study aims to explore the role of high-mobility group box 1 (HMGB1) in an acute glaucoma mouse model. An acute glaucoma model was induced by a rapid and substantial increase IOP to 70 mmHg for 60 min via anterior chamber punctured and affused with Balance Salt Solution in C57BL/6 mice. Retinal tissue ischemic damage and loss of RGCs were assessed at 6, 24, 48, 72 h after high IOP treatment, and at 48 h, group with or without recombinant high-mobility group box 1 (rHMGB1), the HMGB1 inhibitor, glycyrrhizic acid (GA), and by HE and immunofluorescent staining. The nuclear factor κB (NF-κB) inhibitor, JSH-23, and caspase-8 inhibitor, Z-IETD-fmk, were injected into vitreous. Reverse transcription and semi-quantitative reverse transcription polymerase chain reaction (RT-PCR), western blotting, and immunoprecipitation were performed to evaluate the expression level of nucleotide-binding domain, leucine-rich repeat containing protein 3 (NLRP3), phosphor-NF-κB p65, caspase-8, caspase-1, apoptosis-associated speck-like protein containing a CARD (ASC), and interleukin-1β (IL-1β). HMGB1 was increased in ischemic retinal tissue during acute glaucoma as early as 6 h after rapid IOP elevation. Exogenous HMGB1 exacerbated retinal ischemic damage, RGC loss, and inhibition of endogenous HMGB1 significantly reduced the severity of disease. HMGB1 significantly induced the elevation of canonical NLRP3, ASC, caspase-1, and non-canonical capase-8-ASC inflammasome and promoted the processing of IL-1β. Furthermore, the effect of HMGB1 on NLRP3 inflammasome activation and IL-1β production was dependent on NF-κB pathway. Thus, HMGB1/caspase-8 pathway promoted the processing of IL-1β via NF-κB pathway. The findings of this study identified a novel signaling pathway in which HMGB1, in response to acutely elevated intraocular pressure, activated the canonical NLRP3 and non-canonical caspase-8 inflammasomes and production of IL-1β during acute glaucoma development. These results provide new insights to the understanding of the innate response that contributes to pathogenesis of acute glaucoma.