Ng V, Cho P, To C
Department of Optometry & Radiography, The Hong Kong Polytechnic University, Kowloon, SAR, China.
Graefes Arch Clin Exp Ophthalmol. 2000 Sep;238(9):738-45. doi: 10.1007/s004170000140.
Analysis of tear proteins is of diagnostic value for abnormal ocular conditions such as dry eye syndrome. Many studies of tear proteins have been performed on Caucasian subjects. However, little is known about these proteins in Chinese eyes.
The total tear protein concentrations of 30 normal young Hong Kong Chinese were determined by the Bradford method and the modified Lowry method. Bovine serum albumin (BSA) and bovine immunoglobulin G (IgG) were both used as standards for each method. The tear protein patterns were determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and the concentrations of major tear proteins were quantified by scanning densitometry after SDS-PAGE.
The mean +/- SD total tear protein concentrations determined by the Bradford method, using BSA and IgG as standards, were 6.05 +/- 1.58 mg/ml and 11.48 +/- 2.32 mg/ml respectively. The values determined by the modified Lowry method, using the same two standards, were 9.66 +/- 2.03 mg/ml and 7.53 +/- 1.80 mg/ml respectively. The mean +/- SD concentrations of major tear proteins were 2.73 +/- 0.82 mg/ml for lactoferrin, 0.021 +/- 0.028 mg/ml for human serum albumin, 2.89 +/- 0.88 mg/ml for tear-specific prealbumin and 2.46 +/- 0.44 mg/ml for lysozyme.
The results of total tear protein concentrations indicated that values obtained from different methods and different standards were not comparable. The tear protein patterns of our subjects were qualitatively similar to those reported for Caucasian subjects. However, the concentrations of the major proteins of our subjects were not in accordance with those reported previously. The main reason may be the large variability of method used.
泪液蛋白质分析对于诸如干眼症等异常眼部状况具有诊断价值。许多关于泪液蛋白质的研究是在白种人受试者身上进行的。然而,对于中国人眼中的这些蛋白质知之甚少。
采用考马斯亮蓝法和改良的洛里法测定30名正常香港年轻中国人的总泪液蛋白质浓度。每种方法均使用牛血清白蛋白(BSA)和牛免疫球蛋白G(IgG)作为标准品。通过十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳(SDS - PAGE)确定泪液蛋白质图谱,并在SDS - PAGE后通过扫描密度计对主要泪液蛋白质的浓度进行定量。
以BSA为标准品,考马斯亮蓝法测定的总泪液蛋白质平均浓度±标准差为6.05±1.58mg/ml;以IgG为标准品时,该值为11.48±2.32mg/ml。使用相同的两种标准品,改良洛里法测定的值分别为9.66±2.03mg/ml和7.53±1.80mg/ml。主要泪液蛋白质的平均浓度±标准差分别为:乳铁蛋白2.73±0.82mg/ml,人血清白蛋白0.021±0.028mg/ml,泪液特异性前白蛋白2.89±0.88mg/ml,溶菌酶2.46±0.44mg/ml。
总泪液蛋白质浓度的结果表明,不同方法和不同标准获得的值不可比。我们研究对象的泪液蛋白质图谱在质量上与白种人受试者的报道相似。然而,我们研究对象主要蛋白质的浓度与先前报道的不一致。主要原因可能是所使用方法的巨大变异性。