PDF(Pubmed)
Abstract：
Previous studies reported the expression of toll-like receptors (TLRs), merely TLR2 and TLR4, and complement fragments (C3a, C5b9) in vitreoretinal disorders. Other than pathogens, TLRs can recognize endogenous products of tissue remodeling as damage-associated molecular pattern (DAMPs). The aim of this study was to confirm the expression of TLR2 and TLR4 in the fibrocellular membranes and vitreal fluids (soluble TLRs) of patients suffering of epiretinal membranes (ERMs) and assess their association with disease severity, complement fragments and inflammatory profiles. Twenty (n = 20) ERMs and twelve (n = 12) vitreous samples were collected at the time of the vitrectomy. Different severity-staged ERMs were processed for: immunolocalization (IF), transcriptomic (RT-PCR) and proteomics (ELISA, IP/WB, Protein Chip Array) analysis. The investigation of targets included TLR2, TLR4, C3a, C5b9, a few selected inflammatory biomarkers (Eotaxin-2, Rantes, Vascular Endothelial Growth Factor (VEGFA), Vascular Endothelial Growth Factor receptor (VEGFR2), Interferon-γ (IFNγ), Interleukin (IL1β, IL12p40/p70)) and a restricted panel of matrix enzymes (Matrix metalloproteinases (MMPs)/Tissue Inhibitor of Metallo-Proteinases (TIMPs)). A reduced cellularity was observed as function of ERM severity. TLR2, TLR4 and myD88 transcripts/proteins were detected in membranes and decreased upon disease severity. The levels of soluble TLR2 and TLR4, as well as C3a, C5b9, Eotaxin-2, Rantes, VEGFA, VEGFR2, IFNγ, IL1β, IL12p40/p70, MMP7 and TIMP2 levels were changed in vitreal samples. Significant correlations were observed between TLRs and complement fragments and between TLRs and some inflammatory mediators. Our findings pointed at TLR2 and TLR4 over-expression at early stages of ERM formation, suggesting the participation of the local immune response in the severity of disease. These activations at the early-stage of ERM formation suggest a potential persistence of innate immune response in the early phases of fibrocellular membrane formation.
摘要：
先前的研究报道了toll样受体（TLRs）的表达，仅TLR2和TLR4，以及补体片段(C3a，C5b9）在玻璃体视网膜疾病中。除了病原体，TLR可以将组织重塑的内源性产物识别为损伤相关分子模式(DAMPs)。这项研究的目的是确认TLR2和TLR4在患有视网膜前膜（ERMs）的患者的纤维细胞膜和玻璃体液体（可溶性TLRs）中的表达，并评估其与疾病严重程度的关系。补体片段和炎症特征。在玻璃体切除术时收集20个(n=20)ERM和12个(n=12)玻璃体样品。对不同严重程度分期的ERM进行了处理：免疫定位（IF），转录组学(RT-PCR)和蛋白质组学(ELISA,IP/WB,蛋白芯片阵列)分析。调查的目标包括TLR2，TLR4，C3a，C5b9，一些选定的炎症生物标志物(Eotaxin-2，Rantes，血管内皮生长因子(VEGFA),血管内皮生长因子受体(VEGFR2),干扰素-γ(IFNγ),白细胞介素（IL1β，IL12p40/p70））和一组受限的基质酶（基质金属蛋白酶（MMP）/金属蛋白酶组织抑制剂（TIMPs）。观察到作为ERM严重程度的函数的细胞减少。在膜中检测到TLR2、TLR4和myD88转录物/蛋白,并且随着疾病严重程度而降低。可溶性TLR2和TLR4，以及C3a的水平，C5b9Eotaxin-2Rantes,VEGFA,VEGFR2,IFNγ,IL1β,在玻璃体样品中IL12p40/p70、MMP7和TIMP2水平改变。在TLR和补体片段之间以及在TLR和一些炎症介质之间观察到显著的相关性。我们的发现指出TLR2和TLR4在ERM形成的早期过度表达。提示局部免疫反应参与疾病的严重程度。在ERM形成的早期阶段的这些激活表明在纤维细胞膜形成的早期阶段先天性免疫应答的潜在持久性。
