Solomon A, Rosenblatt M, Monroy D, Ji Z, Pflugfelder S C, Tseng S C
Ocular Surface and Tear Center, Bascom Palmer Eye Institute, Department of Ophthalmology, University of Miami School of Medicine, Miami, Florida, USA.
Br J Ophthalmol. 2001 Apr;85(4):444-9. doi: 10.1136/bjo.85.4.444.
Amniotic membrane (AM) transplantation reduces inflammation in a variety of ocular surface disorders. The aim of this study was to determine if AM stroma suppresses the expression of the IL-1 gene family in cultured human corneal limbal epithelial cells.
Human corneal limbal epithelial cells were cultured from limbocorneal explants of donor eyes on plastic or on the AM stroma. Transcript expression of IL-1alpha, IL-1beta, IL-1 receptor antagonist (RA), and GAPDH was compared with or without addition of lipopolysaccharide to their serum-free media for 24 hours using RNAse protection assay (RPA). Their protein production in the supernatant was analysed by ELISA.
Expression of IL-1alpha and IL-1beta transcripts and proteins was significantly reduced by cells cultured on the AM stromal matrix compared with plastic cultures whether lipopolysaccharide was added or not. Moreover, expression of IL-1 RA by cells cultured in the lipopolysaccharide-free medium was upregulated by AM stromal matrix. The ratio between IL-1 RA and IL-1alpha protein levels in AM cultures was higher than in plastic cultures.
AM stromal matrix markedly suppresses lipopolysaccharide induced upregulation of both IL-1alpha and IL-1beta. These data may explain in part the effect of AM transplantation in reducing ocular surface inflammation, underscoring the unique feature of the AM as a substrate for tissue engineering.
羊膜移植可减轻多种眼表疾病中的炎症。本研究旨在确定羊膜基质是否能抑制培养的人角膜缘上皮细胞中白细胞介素 -1（IL-1）基因家族的表达。
从供体眼的角膜缘外植体中培养人角膜缘上皮细胞，分别培养于塑料培养皿或羊膜基质上。使用核糖核酸酶保护分析（RPA），比较在无血清培养基中添加或不添加脂多糖培养24小时后，IL-1α、IL-1β、IL-1受体拮抗剂（RA）和甘油醛 -3-磷酸脱氢酶（GAPDH）的转录本表达情况。通过酶联免疫吸附测定（ELISA）分析其在上清液中的蛋白质生成情况。
无论是否添加脂多糖，与塑料培养相比，在羊膜基质上培养的细胞中IL-1α和IL-1β转录本及蛋白质的表达均显著降低。此外，在无脂多糖培养基中培养的细胞，其IL-1 RA的表达被羊膜基质上调。羊膜培养中IL-1 RA与IL-1α蛋白水平的比值高于塑料培养。
羊膜基质显著抑制脂多糖诱导的IL-1α和IL-1β上调。这些数据可能部分解释了羊膜移植在减轻眼表炎症方面的作用，突出了羊膜作为组织工程基质的独特特性。
