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花粉酶可降解人泪液和结膜细胞:一种理解季节性非过敏性结膜炎的方法。

Pollen enzymes degrade human tear fluid and conjunctival cells: an approach to understanding seasonal non-allergic conjunctivitis.

作者信息

Rabensteiner Dieter Franz, Spreitzhofer Eva, Trummer Gabriele, Wachswender Christine, Kirchengast Sieglinde, Horwath-Winter Jutta, Schmut Otto

出版信息

Dev Ophthalmol. 2010;45:83-92. doi: 10.1159/000315022. Epub 2010 May 18.

Abstract

BACKGROUND

During pollen seasons, allergy-like symptoms can be observed in proven non-allergy sufferers. Pollen enzymes are thought to be responsible for conjunctival irritation. We investigated the influence of the well-known aggressive pollen species hazelnut (Corylus avellana) and birch pollen (Betula pendula) on both human tear fluid and conjunctival cell cultures. This study is an approach to seasonal non-allergic conjunctivitis (SNAC) syndrome.

METHODS

Zymography was carried out in order to investigate the proteolytic activity of the pollen. Thereafter, human tear fluid was incubated with pollen extract, and the results were studied by polyacrylamide gel electrophoresis. In addition, cultivated conjunctival cells (CHANG cells) were incubated with pollen extracts. Cytomorphological changes were analyzed using the CASY1 Cell Counter. Cell viability was quantified via MTS assay. The viability of the cells which were incubated with pollen extract was compared to the viability of control cells.

RESULTS

Pollen proteases destroy tear fluid proteins, as observed by polyacrylamide gel electrophoresis. The treatment of CHANG cells with pollen extract induced a statistically significant decrease in cell viability, depending on the pollen extract concentration and the incubation period.

CONCLUSION

Evidence of the destruction of tear fluid proteins and damage to human conjunctival cells by pollen proteases explains conjunctival irritation in proven non-allergic people during the pollen season. One reason why not all people are affected by SNAC syndrome to the same extent could be differences in the concentrations of antiproteases present on the ocular surface.

摘要

背景

在花粉季节,已证实的非过敏患者也会出现类似过敏的症状。花粉酶被认为是导致结膜刺激的原因。我们研究了著名的具有侵袭性的花粉种类榛树花粉(欧洲榛)和桦树花粉(垂枝桦)对人泪液和结膜细胞培养物的影响。本研究是针对季节性非过敏性结膜炎(SNAC)综合征的一种研究方法。

方法

进行酶谱分析以研究花粉的蛋白水解活性。此后,将人泪液与花粉提取物一起孵育,并通过聚丙烯酰胺凝胶电泳研究结果。此外,将培养的结膜细胞(CHANG细胞)与花粉提取物一起孵育。使用CASY1细胞计数器分析细胞形态学变化。通过MTS测定法定量细胞活力。将与花粉提取物孵育的细胞的活力与对照细胞的活力进行比较。

结果

通过聚丙烯酰胺凝胶电泳观察到,花粉蛋白酶会破坏泪液蛋白。用花粉提取物处理CHANG细胞会导致细胞活力在统计学上显著下降,这取决于花粉提取物的浓度和孵育时间。

结论

花粉蛋白酶破坏泪液蛋白并损害人结膜细胞的证据解释了在花粉季节已证实的非过敏人群中结膜刺激的现象。并非所有人都受到SNAC综合征同等程度影响的一个原因可能是眼表存在的抗蛋白酶浓度不同。

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