Chen Ziyan, Shamsi Farrukh A, Li Kaijun, Huang Qiang, Al-Rajhi Ali A, Chaudhry Imtiaz A, Wu Kaili
Zhongshan Ophthalmic Center, State Key Laboratory of Ophthalmology, Sun Yat-sen University, Guangzhou, China.
Mol Vis. 2011 Feb 1;17:323-31.
Proteins in the tear fluid have positive effects on maintaining the integrity and stabilization of the tear film, which is affected by several environmental factors. The aim of this study is to investigate seasonal variation of protein patterns in camel tears collected during the summer and winter season.
Tears from both eyes of 50 clinically normal camels (Camelus dromedarius) were collected in the summer (June-July) and in the winter (December-January) respectively. Pooled tear protein samples from two seasons were separated by SDS-PAGE and two-dimensional electrophoresis (2-DE). Protein spots of differential expression in two season gels were excised and subjected to in-gel digestion and identification by matrix assisted laser desorption/ionization-time of flight/time of flight-mass spectrum (MALDI-TOF/TOF-MS) analysis. Two differentially expressed proteins, lactoferrin (LF) and vitelline membrane outer layer protein 1 homolog (VMO1 homolog), were validated by western blotting.
Thirteen well resolved bands were detected in SDS-PAGE gels of both summer and winter camel tears. By band densitometry, significantly higher intensities of band 6, 7, 11, and lower intensity of band 13 were observed in the summer group compared to the winter group. In 2-DE profiles of camel tears, four protein spots were found expressed differentially in two seasons. Further protein identification by MALDI-TOF/TOF-MS and confirmation by western blotting indicated that there was a significant decrease in LF (p=0.002) and an increase in VMO1 homolog (p=0.042) in tears in the summer compared to the winter.
The seasonal variation of camel tear fluids has been found in the composition of proteins, including LF and VMO1 homolog. This result will expand our knowledge of physiologic characteristics of tear fluids and establish a foundation for the mechanistic studies and clinical practices on ocular surface disorders.
泪液中的蛋白质对维持泪膜的完整性和稳定性具有积极作用,而泪膜会受到多种环境因素的影响。本研究旨在调查夏季和冬季采集的骆驼泪液中蛋白质模式的季节性变化。
分别于夏季(6月至7月)和冬季(12月至1月)采集50头临床正常单峰骆驼(骆驼属双峰驼)双眼的泪液。将两个季节的混合泪液蛋白质样本通过SDS-PAGE和二维电泳(2-DE)进行分离。切除两个季节凝胶中差异表达的蛋白质斑点,进行胶内消化,并通过基质辅助激光解吸/电离飞行时间/飞行时间质谱(MALDI-TOF/TOF-MS)分析进行鉴定。通过蛋白质印迹法验证了两种差异表达的蛋白质,即乳铁蛋白(LF)和卵黄膜外层蛋白1同源物(VMO1同源物)。
在夏季和冬季骆驼泪液的SDS-PAGE凝胶中均检测到13条清晰分辨的条带。通过条带密度测定法,与冬季组相比,夏季组中条带6、7、11的强度明显更高,条带13的强度更低。在骆驼泪液的2-DE图谱中,发现有4个蛋白质斑点在两个季节中差异表达。通过MALDI-TOF/TOF-MS进一步进行蛋白质鉴定并通过蛋白质印迹法进行确认,结果表明与冬季相比,夏季泪液中LF显著减少(p=0.002),VMO1同源物增加(p=0.042)。
已发现骆驼泪液在蛋白质组成方面存在季节性变化,包括LF和VMO1同源物。这一结果将扩展我们对泪液生理特征的认识,并为眼表疾病的机制研究和临床实践奠定基础。