Retinal Pigment Epithelium Cell Death Is Associated With NLRP3 Inflammasome Activation by All-trans Retinal
Retinal Pigment Epithelium Cell Death Is Associated With NLRP3 Inflammasome Activation by All-trans Retinal
复制标题
视网膜色素上皮细胞死亡与全反式视网膜激活 NLRP3 炎症小体相关
DOI：
10.1167/iovs.18-26360
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发表时间：
2019-07-01
影响因子：
4.4
通讯作者：
Wu, Yalin
中科院分区：
文献类型：
--
作者：
Liao, Yi;Zhang, Houjian;Wu, Yalin
目的.视觉（类维生素A）周期异常诱导视网膜色素上皮（RPE）中全反式视网膜（atRAL）的异常积聚，这是1型Stargardt病和年龄相关性黄斑变性中RPE萎缩的原因。NLR家族pyrin domain containing 3（NLRP 3）炎性小体激活与年龄相关性黄斑变性的病因学有关。在此，我们阐明了NLRP 3炎性小体激活与atRAL诱导的RPE细胞死亡之间的关系。通过MTS或MTT测定评估细胞毒性。mRNA和蛋白质的表达水平通过定量逆转录-聚合酶链反应、蛋白质印迹或酶联免疫吸附测定来确定。使用荧光显微镜检查细胞内信号。透射电子显微镜观察细胞器的超微结构特征。atRAL的异常积累与表现出凋亡和Caspase-3/gasdermin E（GSDME）介导的焦亡特征的人ARPE-19细胞比例的显著增加相关。这些细胞还表现出NLRP 3、ASC、裂解的半胱天冬酶-1/聚ADP-核糖聚合酶（PARP）/半胱天冬酶-3/GSDME、白细胞介素-1 β（IL-1 β）和IL-18以及NLRP 3炎性小体相关基因（IL 1B和IL 18）的表达升高。在人ARPE-19细胞暴露于过量atRAL后，从溶酶体释放的活性氧（ROS）（包括线粒体ROS）和组织蛋白酶传递信号，导致NLRP 3炎性小体活化。抑制ROS、NLRP 3炎性体、Caspase-1、组织蛋白酶B或组织蛋白酶D的产生保护ARPE-19细胞免受atRAL相关的细胞毒性。在atRAL暴露的ARPE-19细胞中，线粒体、溶酶体和内质网的损伤通过用选择性NLRP 3炎性小体转运蛋白MCC 950处理而部分减轻。atRAL的异常积聚通过NLRP 3炎性小体激活促进RPE细胞的死亡。
PURPOSE. Visual (retinoid) cycle anomalies induce aberrant build-up of all-trans retinal (atRAL) in the retinal pigment epithelium (RPE), which is a cause of RPE atrophy in Stargardt disease type 1 and age-related macular degeneration. NLR family pyrin domain containing 3 (NLRP3) inflammasome activation is implicated in the etiology of age-related macular degeneration. Here, we elucidated the relationship between NLRP3 inflammasome activation and atRAL-induced death of RPE cells.METHODS. Cellular toxicities were assessed by MTS or MTT assays. Expression levels of mRNAs and proteins were determined by quantitative reverse transcription-polymerase chain reaction, Western blotting, or enzyme-linked immunosorbent assay. Fluorescence microscopy was used to examine intracellular signals. Ultrastructural features of organelles were examined by transmission electron microscope.RESULTS. Abnormal accumulation of atRAL was associated with a significant increase in the proportion of human ARPE-19 cells exhibiting features of apoptosis and Caspase-3/gasdermin E (GSDME)-mediated pyroptosis. These cells also exhibited elevated expression of NLRP3, ASC, cleaved Caspase-1/poly ADP-ribose polymerase (PARP)/Caspase-3/GSDME, interleukin-1 beta (IL-1 beta), and IL-18, as well as NLRP3 inflammasome-related genes (IL1B and IL18). After exposure of human ARPE-19 cells to excess atRAL, reactive oxygen species (ROS) (including mitochondrial ROS) and cathepsins released from lysosomes transmitted signals leading to NLRP3 inflammasome activation. Suppressing the production of ROS, NLRP3 inflammasome, Caspase-1, cathepsin B, or cathepsin D protected ARPE-19 cells against atRAL-associated cytotoxicity. Damage to mitochondria, lysosomes, and endoplasmic reticulum in atRAL-exposed ARPE-19 cells was partially alleviated by treatment with MCC950, a selective NLRP3 inflammasome inhibitor.CONCLUSIONS. Aberrant build-up of atRAL promotes the death of RPE cells via NLRP3 inflammasome activation.