Elevated inflammatory cytokines contribute to the pathogenesis of various retinal diseases such as diabetic retinopathy, retinal vasculitis and retinitis. However, the underlying mechanism of retinal inflammation remains largely unknown. Recent studies demonstrated that acetylcholinesterase (ACHE) is an inflammatory indicator in central neural system. This study was aimed to dissect the role of ACHE in retinal inflammation, and its mechanism of action. Retinal inflammation was induced by intravitreal injection of tumor necrosis factor-α (TNF-α) in heterozygous ACHE knockdown mice (ACHE+/-) and wild type mice (ACHE+/+). Donepezil, a well-known ACHE inhibitor, was administrated by daily gavage. Expression of ACHE and intercellular adherent molecule-1 (ICAM-1), infiltration of CD11b+ inflammatory cells, retinal leukostasis and vascular leakage was determined in both ACHE+/- and ACHE+/+ mice. ARPE-19 cells, a human retinal pigment epithelial cell line, were cultured for in vitro assay. Knockdown of ACHE was achieved by lipofectamine-mediated siRNA transfection and pharmaceutical suppression of ACHE was manipulated by donepezil. Cellular expression and distribution of ACHE, ICAM-1, and phosphorylation of NF-κB, IκB and IKKα/β were detected by western-blot analysis or immunocytochemistry. Retinal expression of ACHE was dramatically upregulated, in parallel with increased ICAM-1 expression, enhanced leukostasis and augmented CD11b+ inflammatory cell infiltration as well as vascular hyperpermeability in ACHE+/+ mice injected with TNF-α. However, TNF-α-injected ACHE+/- mice showed lower level of ICAM-1, less leukostasis and fewer infiltrated CD11b+ cells. Moreover, TNF-α-induced retinal vascular leakage was significantly reduced in ACHE+/- mice. Similarly, TNF-α-induced retinal inflammatory response were also attenuated by donepezil intervention. In addition, TNF-α treatment resulted in significant induction of ACHE, upregulation of ICAM-1 and nuclear translocation of NF-κB, phosphorylation of IκB and IKKα/β in ARPE-19 cells. However, inhibition of ACHE reduced TNF-α-induced phosphorylation of NF-κB, IκB and IKKα/β in ARPE-19 cells. The present study reveals a pivotal role of ACHE in retinal inflammation. Inhibition of ACHE attenuates retinal inflammation and retinal leakage likely through suppressing NF-κB signaling activation.

中文翻译：

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抑制乙酰胆碱酯酶可通过抑制NF-κB活化来减轻视网膜炎症。

升高的炎性细胞因子促成各种视网膜疾病例如糖尿病性视网膜病，视网膜血管炎和视网膜炎的发病机理。然而，视网膜炎症的潜在机制仍是未知之数。最近的研究表明，乙酰胆碱酯酶（ACHE）是中枢神经系统的炎症指标。这项研究旨在剖析ACHE在视网膜炎症中的作用及其作用机理。在杂合性ACHE敲除小鼠（ACHE +/-）和野生型小鼠（ACHE + / +）中，玻璃体内注射肿瘤坏死因子-α（TNF-α）诱发视网膜炎症。多奈哌齐是一种著名的ACHE抑制剂，每天通过管饲法给药。ACHE和细胞间黏附分子-1（ICAM-1）的表达，CD11b +炎性细胞的浸润，在ACHE +/-和ACHE + / +小鼠中都测定了视网膜白细胞停滞和血管渗漏。培养ARPE-19细胞（一种人类视网膜色素上皮细胞系）进行体外测定。通过脂转染胺介导的siRNA转染来实现ACHE的降低，而多奈哌齐可控制ACHE的药物抑制作用。通过蛋白质印迹分析或免疫细胞化学检测ACHE，ICAM-1的细胞表达和分布以及NF-κB，IκB和IKKα/β的磷酸化。在注射TNF-α的ACHE + / +小鼠中，ACHE的视网膜表达显着上调，同时增加了ICAM-1表达，白细胞增多和CD11b +炎性细胞浸润以及血管通透性增加。但是，注射TNF-α的ACHE +/-小鼠显示出较低的ICAM-1水平，白细胞减少和CD11b +浸润细胞减少。此外，在ACHE +/-小鼠中，TNF-α诱导的视网膜血管渗漏明显减少。同样，多奈哌齐干预也减弱了TNF-α诱导的视网膜炎症反应。此外，TNF-α处理导致ARPE-19细胞中ACHE明显诱导，ICAM-1上调和NF-κB核易位，IκB和IKKα/β磷酸化。但是，抑制ACHE可以减少TNF-α诱导的ARPE-19细胞中NF-κB，IκB和IKKα/β的磷酸化。本研究揭示了ACHE在视网膜炎症中的关键作用。抑制ACHE可能通过抑制NF-κB信号传导激活来减轻视网膜炎症和视网膜渗漏。多奈哌齐干预也减弱了TNF-α诱导的视网膜炎症反应。此外，TNF-α处理导致ARPE-19细胞中ACHE明显诱导，ICAM-1上调和NF-κB核易位，IκB和IKKα/β磷酸化。但是，抑制ACHE可以减少TNF-α诱导的ARPE-19细胞中NF-κB，IκB和IKKα/β的磷酸化。本研究揭示了ACHE在视网膜炎症中的关键作用。抑制ACHE可能通过抑制NF-κB信号传导激活来减轻视网膜炎症和视网膜渗漏。多奈哌齐干预也减弱了TNF-α诱导的视网膜炎症反应。此外，TNF-α处理导致ARPE-19细胞中ACHE明显诱导，ICAM-1上调和NF-κB核易位，IκB和IKKα/β磷酸化。但是，抑制ACHE可以减少TNF-α诱导的ARPE-19细胞中NF-κB，IκB和IKKα/β的磷酸化。本研究揭示了ACHE在视网膜炎症中的关键作用。抑制ACHE可能通过抑制NF-κB信号传导激活来减轻视网膜炎症和视网膜渗漏。抑制ACHE可以减少TNF-α诱导的ARPE-19细胞中NF-κB，IκB和IKKα/β的磷酸化。本研究揭示了ACHE在视网膜炎症中的关键作用。抑制ACHE可能通过抑制NF-κB信号传导激活来减轻视网膜炎症和视网膜渗漏。抑制ACHE可以减少TNF-α诱导的ARPE-19细胞中NF-κB，IκB和IKKα/β的磷酸化。本研究揭示了ACHE在视网膜炎症中的关键作用。抑制ACHE可能通过抑制NF-κB信号传导激活来减轻视网膜炎症和视网膜渗漏。