Hori Yuichi, Argüeso Pablo, Spurr-Michaud Sandra, Gipson Ilene K
Schepens Eye Research Institute, Department of Ophthalmology, Harvard Medical School, Boston, MA 02114, USA.
Cornea. 2006 Feb;25(2):176-81. doi: 10.1097/01.ico.0000177838.38873.2f.
This study was designed to determine whether long-term tolerant contact lens (CL) wear causes changes in the expression of mucin mRNA by the conjunctival epithelium and mucin protein content in tears and to determine whether specific mucins adhere to contact lenses.
Twenty long-term (> or = 5 years ) and tolerant CL wearers (2 with hard and 18 with soft contact lenses) were compared with 23 non-CL wearers. One hour after CL removal, tear fluid was collected after instillation of 60 microL of sterile water onto the ocular surface, and protein concentration was determined. Impression cytology was performed on the bulbar temporal region of conjunctiva to collect cells for RNA isolation. Real-time polymerase chain reaction was performed using TaqMan primer and probes for MUC1, 4, 5AC, and 16. ELISA was performed on the collected tears to detect MUC5AC and the mucin carbohydrate epitope H185. For the analysis of adherent mucins on CL, discarded daily-wear contact lenses were collected, rinsed, and incubated overnight at 4 degrees C in mucin isolation buffer. Immunoblot analysis of adherent mucins was performed to detect MUC1, 4, 5AC, 16, and H185.
No significant changes in the levels of mucin mRNA from impression cytology samples were detected when comparing CL and non-CL wearers. The amount of total protein in tears collected from CL wearers (39.9 +/- 27.2 microg) was significantly less than that from non-CL wearers (95.1 +/- 73.8 microg, P = 0.001). The level of MUC5AC mucin and the H185 epitope in tears per unit protein in CL wearers was not significantly different from non-CL wearers. Low levels of membrane-associated mucins, the secreted mucin MUC5AC, and the carbohydrate epitope, H185, were detected in protein extracts from discarded CLs. Compared with MUC1, 4, and 5AC, there was less MUC16 adherent to the CLs.
Neither mucin mRNA expression by conjunctival epithelia nor mucin content per unit protein in tears was altered by long-term tolerant CL wear; however, the amount of protein in the tears was significantly less. Shed membrane-associated mucins and the goblet cell mucins adhere to CLs.
本研究旨在确定长期耐受佩戴隐形眼镜(CL)是否会导致结膜上皮粘蛋白mRNA表达以及泪液中粘蛋白含量发生变化,并确定特定粘蛋白是否会附着在隐形眼镜上。
将20名长期(≥5年)耐受佩戴隐形眼镜者(2名佩戴硬性隐形眼镜,18名佩戴软性隐形眼镜)与23名非隐形眼镜佩戴者进行比较。取下隐形眼镜1小时后,在眼表滴入60微升无菌水后收集泪液,并测定蛋白质浓度。在结膜颞侧球结膜区域进行印迹细胞学检查,收集细胞用于RNA分离。使用针对MUC1、4、5AC和16的TaqMan引物和探针进行实时聚合酶链反应。对收集的泪液进行酶联免疫吸附测定(ELISA),以检测MUC5AC和粘蛋白碳水化合物表位H185。为了分析隐形眼镜上附着的粘蛋白,收集丢弃的日戴型隐形眼镜,冲洗后在4℃下于粘蛋白分离缓冲液中孵育过夜。对附着的粘蛋白进行免疫印迹分析,以检测MUC1、4、5AC、16和H185。
比较隐形眼镜佩戴者和非隐形眼镜佩戴者时,未检测到印迹细胞学样本中粘蛋白mRNA水平有显著变化。从隐形眼镜佩戴者收集的泪液中总蛋白量(39.9±27.2微克)明显低于非隐形眼镜佩戴者(95.1±73.8微克,P = 0.001)。隐形眼镜佩戴者泪液中每单位蛋白的MUC5AC粘蛋白水平和H185表位与非隐形眼镜佩戴者无显著差异。在丢弃的隐形眼镜的蛋白质提取物中检测到低水平的膜相关粘蛋白、分泌型粘蛋白MUC5AC和碳水化合物表位H185。与MUC1、4和5AC相比,附着在隐形眼镜上的MUC16较少。
长期耐受佩戴隐形眼镜既不会改变结膜上皮的粘蛋白mRNA表达,也不会改变泪液中每单位蛋白的粘蛋白含量;然而,泪液中的蛋白量明显较少。脱落的膜相关粘蛋白和杯状细胞粘蛋白会附着在隐形眼镜上。