Sitaramamma T, Shivaji S, Rao G N
Prof. Brien Holden Eye Research Centre, L.V. Prasad Eye Institute, Hyderabad, India.
Indian J Ophthalmol. 1998 Dec;46(4):239-45.
Changes in the closed, open and reflex eye tear proteins of normal subjects were compared and analysed. Tear proteins were resolved by high-performance liquid chromatography (HPLC) utilising both gel filtration (P-300 SW) and reverse-phase (C-18) columns and the HPLC fractions were further analysed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) under reducing and non-reducing conditions. The protein composition of the closed-eye tear was significantly different from that of the open and reflex-eye tear. Secretory IgA (sIgA) was the predominant protein in closed eye tears constituting 49% of the total protein compared to 11% in reflex tears, whereas lysozyme was the predominant protein (53%) in reflex tears. Levels of lactoferrin, lipocalin and lysozyme were relatively constant in both open and reflex tears. HPLC profiles of the closed-eye tears, upon continuous stimulation of lacrimal glands indicated that sIgA was significantly reduced whereas lactoferrin, lipocalin, and lysozyme were significantly increased. These results indicate that the tear composition upon waking attains that of the open eye within 4 to 5 minutes, and upon continuous stimulation this reflects the reflex-eye tear composition. It also indicates that mechanisms responsible for changes in concentration of constitutive and regulated tear protein with stimulus can be studied successfully using non-invasive methods to collect human tears.
对正常受试者闭眼、睁眼和反射性流泪时的泪液蛋白质变化进行了比较和分析。利用凝胶过滤柱（P - 300 SW）和反相柱（C - 18）通过高效液相色谱（HPLC）分离泪液蛋白质，并且在还原和非还原条件下通过十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳（SDS - PAGE）对HPLC馏分进行进一步分析。闭眼时泪液的蛋白质组成与睁眼和反射性流泪时的蛋白质组成显著不同。分泌型IgA（sIgA）是闭眼时泪液中的主要蛋白质，占总蛋白质的49%，而在反射性泪液中占11%，而溶菌酶是反射性泪液中的主要蛋白质（53%）。乳铁蛋白、脂钙蛋白和溶菌酶的水平在睁眼和反射性泪液中相对恒定。在持续刺激泪腺时，闭眼泪液的HPLC图谱表明sIgA显著减少，而乳铁蛋白、脂钙蛋白和溶菌酶显著增加。这些结果表明，醒来时的泪液成分在4至5分钟内达到睁眼时的成分，并且在持续刺激时反映反射性泪液的成分。这也表明，使用非侵入性方法收集人泪液能够成功研究刺激引起的组成性和调节性泪液蛋白质浓度变化的机制。
