Subconjunctival injections of dimethyl fumarate inhibit lymphangiogenesis and allograft rejection in the rat cornea
Subconjunctival injections of dimethyl fumarate inhibit lymphangiogenesis and allograft rejection in the rat cornea
复制标题
结膜下注射富马酸二甲酯抑制大鼠角膜淋巴管生成和同种异体移植排斥
DOI：
10.1016/j.intimp.2021.107580
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发表时间：
2021-04-03
影响因子：
5.6
通讯作者：
Liang，Dan
中科院分区：
文献类型：
--
作者：
Yu，Jianfeng;Li，Yingqi;Liang，Dan
巨噬细胞诱导的角膜淋巴管生成在角膜异体移植排斥反应（CGR）中起关键作用。然而，很少有食品和药物管理局（FDA）批准的针对淋巴管生成的药物。本研究旨在探讨富马酸二甲酯（DMF）对大鼠同种异体角膜移植存活的影响。大鼠进行穿透性角膜移植。在手术结束和术后第3天至第11天给予富马酸二甲酯（20µg）结膜下注射。观察异体角膜移植的临床表现。免疫组织化学、实时荧光定量PCR （qPCR）、流式细胞术和western blot检测。采用qPCR、酶联免疫吸附试验（ELISA）和体外western blotin检测DMF对RAW264.7细胞的作用及机制。结果显示，与载药组相比，结膜下注射DMF可显著抑制角膜淋巴管生成和CGR，角膜巨噬细胞浸润减少。DMF可通过抑制NF-κB活化，降低角膜移植物和RAW264.7巨噬细胞单核细胞趋化蛋白1 （MCP-1）、肿瘤坏死因子-α (TNF-α)、白细胞介素-6 （IL-6）、白细胞介素-1β (IL-1β)和血管内皮生长因子- c (VEGF-C) mRNA的表达。此外，与载药组相比，dmf处理组同侧颈部淋巴结树突状细胞数量明显减少。综上所述，我们的研究结果表明DMF可以通过抑制巨噬细胞诱导的角膜淋巴血管生成来抑制CGR。
Corneal lymphangiogenesis induced by macrophages played a critical role in corneal allograft rejection (CGR). However, there are few Food and Drug Administration (FDA)-approved drugs that target lymphangiogenesis. The aim of our study is to evaluate the effects of dimethyl fumarate (DMF) on corneal allograft survival in rats. Penetrating corneal transplantation was performed in rats. Subconjunctival injections of dimethyl fumarate (20 µg) were administered at the end of the operation and postoperative day 3 to day 11. The clinical signs of corneal allografts were evaluated. Immunohistochemistry, quantitative real-time PCR (qPCR), flow cytometry and western blot were performed respectively. The effects and mechanism of DMF on RAW264.7 cells were determined by qPCR, enzyme-linked immunosorbent assay (ELISA), and western blotin vitro. The results showed that subconjunctival injections of DMF could significantly inhibit corneal lymphangiogenesis and CGR with decreased corneal macrophage infiltration compared with the vehicle group. Moreover, DMF could reduce the mRNA expression of monocyte chemoattractant protein 1 (MCP-1), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), interleukin-1β (IL-1β), and vascular endothelial growth factor-C (VEGF-C) in the corneal grafts and RAW264.7 macrophages by inhibiting NF-κB activation. Furthermore, compared with the vehicle group, the number of dendritic cells in the ipsilateral cervical lymph nodes of the DMF-treated group was decreased significantly. Collectively, our findings showed that DMF could suppress CGR by inhibiting the macrophage-induced corneal lymphoangiogenesis.