Characterization of human ocular mucin secretion mediated by 15(S)-HETE.
Characterization of human ocular mucin secretion mediated by 15(S)-HETE.
复制标题
15(S)-HETE 介导的人眼粘蛋白分泌的表征。
DOI：
10.1097/00003226-200211000-00018
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发表时间：
2002
期刊：
影响因子：
2.8
通讯作者：
Jumblatt,MarciaM
中科院分区：
文献类型：
--
作者：
Jumblatt,JamesE;Cunningham,LaurenT;Li,Yang;Jumblatt,MarciaM
目的：二十烷类化合物15-(S)-羟基-5，8，11，13-二十碳四烯酸[15(S)-HETE]被报道能刺激呼吸道和眼表上皮细胞产生粘蛋白。本研究旨在评价15(S)-HETE对人结膜细胞分泌特异性粘蛋白的影响。方法：将人球结膜组织切片与15(S)-HETE(1-1000 nm)在37℃孵育30分钟，用针对粘蛋白多肽表位的抗体斑点免疫分析法检测培养上清液中人眼粘蛋白MUC1、MUC2、MUC4和MUC5AC的分泌。结果15(S)-HETE(10−8-10−6 M)以浓度依赖的方式刺激结膜粘蛋白的分泌。10−7 M 15(S)-HETE组总粘蛋白分泌显著增加，10−6 M时最高(比对照组增加50%)。免疫分析结果表明，15(S)-HETE对粘蛋白的分泌有不同程度的刺激作用，对MUC2、MUC4和MUC5Ac的释放无明显影响。结论：15(S)-HETE是人结膜组织中MUC1的选择性分泌物。尽管粘蛋白1分泌的生化机制(S)和细胞来源尚不清楚，但在角膜和结膜上皮细胞中广泛表达的粘蛋白1及其在人泪液中的存在表明，分泌的粘蛋白1可能有助于覆盖和保护眼表的粘蛋白层。
Purpose.The eicosanoid 15-(S)-hydroxy-5, 8, 11, 13-eicosatetraenoic acid [15 (S)-HETE] is reported to stimulate mucin production in both airway and ocular surface epithelia. The current study was undertaken to evaluate the effects of 15 (S)-HETE on secretion of specific ocular mucins by human conjunctiva.Methods.Segments of human bulbar conjunctival tissue were incubated with 15 (S)-HETE (1–1000 nM) for 30 minutes at 37 C. Secretion of human ocular mucins MUC1, MUC2, MUC4, and MUC5AC into the incubation media was measured by dot-blot immunoassay using antibodies directed to unique mucin polypeptide epitopes. Sodium dodecyl sulfate–polyacrylamide gel electrophoresis and Western blotting were used to verify the specificity of anti-mucin antibody binding and to investigate the presence of MUC1 mucin in human tears.Results.15 (S)-HETE (10− 8–10− 6 M) stimulated secretion of conjunctival mucins in a concentration-dependent manner. Significant increases in total mucin secretion were observed at 10− 7 M 15 (S)-HETE with a maximum response (> 50% increase above controls) at 10− 6 M. Results of immunoassays showed that 15 (S)-HETE differentially stimulates secretion of MUC1 mucin with no detectable effects on MUC2, MUC4, or MUC5AC release. Western analysis of tear samples from human volunteers indicated that MUC1 is a component of the preocular tear film.Conclusions.The results demonstrate that 15 (S)-HETE is a selective secretogogue for MUC1 in isolated human conjunctival tissue. Although the biochemical mechanism (s) and cellular origins of MUC1 secretion remain to be established, the ubiquitous expression of MUC1 in corneal and conjunctival epithelia and its presence in human tears suggest that secreted MUC1 may contribute to the mucin layer that coats and protects the ocular surface.