Topical application of culture supernatant from human amniotic epithelial cells suppresses inflammatory reactions in cornea
Topical application of culture supernatant from human amniotic epithelial cells suppresses inflammatory reactions in cornea
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DOI：
10.1016/j.exer.2004.11.018
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发表时间：
2005-05-01
影响因子：
3.4
通讯作者：
Hori, J
中科院分区：
文献类型：
--
作者：
Kamiya, K;Wang, MC;Hori, J
人羊膜上皮细胞（HAEC）可能是可溶性抗炎因子的来源。本研究的目的是确定局部应用HAEC培养上清液对角膜炎症反应的影响。从胎盘中提取HAEC，培养48小时，收集上清。从HAEC提取的条件培养基中含有少量人白细胞介素-1受体拮抗剂（IL-1ra）。在BALB/c小鼠角膜内缝合，诱导角膜新生血管形成。采用浅表烧灼术诱导角膜内抗原呈递细胞（APCs）的募集或活化，而不产生新生血管。每日局部应用HAEC条件培养基、安慰剂或重组人IL-1ra 3次，持续2周。在显微镜下观察缝线诱导角膜新生血管的形成情况，持续8周。2周后采集灼烧角膜，免疫荧光染色和共聚焦显微镜检测MHCⅱ+ apc。采用多探针核糖核酸酶保护实验分析角膜灼烧后炎性细胞因子基因的表达。来自HAEC的条件培养基导致角膜新生血管的严重抑制和上皮中MHC II类+ apc的减少。相比之下，人IL-1ra在抑制角膜炎症反应方面只有轻微的效果。应用条件培养基后，小鼠灼烧角膜中IL-1ra和il -1 β mRNA的表达明显受到抑制。这些结果表明，HAEC是可溶性抗炎因子的来源，并且HAEC的条件培养基中含有除IL-1ra以外的抑制角膜炎症的因子。(c) 2004 Elsevier Ltd.版权所有。
Human amniotic epithelial cells (HAEC) may be a source of soluble anti-inflammatory factors. The purpose of this study is to determine the effect of topically applied HAEC culture supernatant on corneal inflammatory reactions. HAEC were obtained from a placenta and cultured for 48 hr, and the supernatant was collected. The conditioned medium from HAEC contained small amounts of human interleukin-1 receptor antagonist (IL-1ra). Intrastromal sutures were placed in the cornea of BALB/c mice to induce corneal neovascularisation. Superficial cauterisation was applied to induce recruitment or activation of antigen presenting cells (APCs) in the cornea without neovascularisation. HAEC conditioned medium, placebo, or recombinant human IL-1ra was topically applied three times daily for 2 weeks. Suture-induced corneal neovascularisation was evaluated microscopically for 8 weeks. The cauterised corneas were harvested at 2 weeks, and the MHC class II+ APCs were quantified by immunofluorescent staining and confocal microscopy. Inflammatory cytokine gene expression in the cauterised corneas was analyzed by a multiprobe ribonuclease protection assay. Conditioned medium from HAEC led to a profound suppression of corneal neovascularisation and fewer MHC class II+ APCs in the epithelium. In contrast, human IL-1ra was only slightly effective in suppressing corneal inflammatory reactions. mRNA expression of murine IL-1ra and IL-1beta in the cauterised corneas was markedly suppressed after application of the conditioned medium. These results suggest that HAEC are a source of soluble anti-inflammatory factors and that conditioned medium from HAEC contains factors other than IL-1ra that suppress corneal inflammation. (c) 2004 Elsevier Ltd. All rights reserved.