Inhibition of TLR4 alleviates the inflammation and apoptosis of retinal ganglion cells in high glucose
Inhibition of TLR4 alleviates the inflammation and apoptosis of retinal ganglion cells in high glucose
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抑制TLR4减轻高糖条件下视网膜神经节细胞的炎症和凋亡
DOI：
10.1007/s00417-017-3772-0
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发表时间：
2017-11-01
影响因子：
2.7
通讯作者：
Jiang, Shuanghong
中科院分区：
文献类型：
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作者：
Hu, Lili;Yang, Hongxia;Jiang, Shuanghong
目的探讨高糖培养视网膜神经节细胞（RGCs）Toll样受体4（TLR 4）的表达谱、TLR 4对炎症和细胞凋亡的影响及其机制。方法建立高糖培养视网膜神经节细胞（RGCs）模型，采用Brn 3a进行鉴定。将原代培养的RGC分为对照（0 mM）、HG 1（10 mM葡萄糖）、HG 2（20 mM葡萄糖）、HG 3（30 mM葡萄糖）、HG（20 mM葡萄糖）+ TAK-242（1.0 μM）和HG（20 mM葡萄糖）+溶剂（1% DMSO）组。在24 h和48 h通过实时定量PCR、Western blot或ELISA检测TLR 4及其下游信号分子和促炎细胞因子的表达水平。结果高糖（10 mM，20 mM，30 mM）组视网膜节细胞TLR 4 mRNA和蛋白表达水平均升高。与这些发现一致，四种TLR 4下游信号分子（MyD 88、NF-κB、TRAF 6、NLRP 3）和促炎细胞因子（IL-1β、IL-18）在三个高糖组中上调。高糖组RGCs凋亡明显增加。TLR 4拮抗剂TAK-242可抑制高糖组RGCs的炎症和凋亡。结论TLR 4在高糖诱导的RGCs炎症和凋亡中起重要作用。TLR 4可能成为预防DR进展的一个新的潜在药理学靶点。
PurposeTo investigate the expression profiles of Toll-like receptor 4 (TLR4), the effect of TLR4 on inflammation, and apoptosis of retinal ganglion cells (RGCs) cultured in high glucose and the underlying mechanism.MethodsA high-glucose model was established in RGCs isolated from Sprague-Dawley (SD) rats (2–3 days old) and identified with Brn3a. Primary cultured RGCs were divided into control (0 mM), HG1 (10 mM glucose), HG2 (20 mM glucose), HG3 (30 mM glucose), HG (20 mM glucose) + TAK-242 (1.0 μM), and HG (20 mM glucose) + vehicle (1% DMSO) groups. The expression levels of TLR4, its downstream signalling molecules, and pro-inflammatory cytokines were measured by real-time PCR, Western blot or ELISA at 24 h and 48 h. The apoptosis rate of RGCs was measured by flow cytometry.ResultsThe mRNA and protein expression levels of TLR4 were increased in high-glucose groups (10 mM, 20 mM, 30 mM). Consistent with these findings, four TLR4 downstream signalling molecules (MyD88, NF-κB, TRAF6, NLRP3) and pro-inflammatory cytokines (IL-1β, IL-18) were upregulated in the three high-glucose groups. Apoptosis of RGCs was clearly increased in the high-glucose group. The administration of TAK-242, an antagonist of TLR4, inhibited inflammation and apoptosis of RGCs in the high-glucose group.ConclusionOur results demonstrated that TLR4 plays a critical role in the inflammation and apoptosis of RGCs induced by high glucose. TLR4 might become a novel potential pharmacological target for preventing the progression of DR.