干燥综合征兔模型泪腺氯通道的变化。
Changes of chloride channels in the lacrimal glands of a rabbit model of Sjögren syndrome.
机构信息
Department of Cell and Neurobiology, Keck School of Medicine, University of Southern California, Los Angeles, CA 90089-9112, USA.
出版信息
Cornea. 2012 Mar;31(3):273-9. doi: 10.1097/ICO.0b013e3182254b42.
PURPOSE
To test the hypothesis that expressions of Na-K-2Cl cotransporter-1 (NKCC1), cystic fibrosis transmembrane conductance regulator (CFTR), and chloride channel 2 γ subunit (ClC2γ) in the lacrimal glands (LGs) of rabbits with induced autoimmune dacryoadenitis (IAD) are changed.
METHODS
LGs were obtained from adult female rabbits with IAD and age-matched female control rabbits. LGs were processed for laser capture microdissection, real-time reverse transcription-polymerase chain reaction, Western blot, and immunofluorescence.
RESULTS
In rabbits with IAD, messenger RNA (mRNA) abundance and protein expressions of NKCC1 and CFTR from whole LGs were significantly lower than those in controls. mRNA abundance of NKCC1, CFTR, and ClC2γ from rabbits with IAD was significantly different from that in acinar and ductal cells from controls. NKCC1 was localized to the basolateral membranes of all acinar and ductal cells, with weaker staining intensity in ductal cells, and the staining pattern from rabbits with IAD appeared similar to that from controls. CFTR was found as punctate aggregates in the apical cytoplasm of all acinar and ductal cells, with the intensity in ductal cells much stronger and no significant difference between controls and rabbits with IAD. ClC2γ was also localized to the apical cytoplasm as punctate aggregates of all acinar cells but not in ductal cells, and a similar staining pattern was observed in rabbits with IAD compared with control rabbits.
CONCLUSIONS
Our data demonstrated significant changes of mRNA and protein expressions of NKCC1, CFTR, and ClC2γ in rabbits with IAD, suggesting that these changes may contribute to the altered lacrimal secretion, particularly Cl transport, in rabbits with IAD.
目的
验证在诱导自身免疫性干眼病（IAD）的兔泪腺中，钠钾 2 氯共转运蛋白 1（NKCC1）、囊性纤维化跨膜电导调节因子（CFTR）和氯离子通道 2γ 亚基（ClC2γ）的表达发生变化的假说。
方法
从患有 IAD 的成年雌性兔和年龄匹配的雌性对照兔中获取泪腺。对泪腺进行激光捕获显微解剖、实时逆转录聚合酶链反应、Western blot 和免疫荧光分析。
结果
在患有 IAD 的兔中，整个泪腺中 NKCC1 和 CFTR 的信使 RNA（mRNA）丰度和蛋白表达均明显低于对照组。IAD 兔的 NKCC1、CFTR 和 ClC2γ 的 mRNA 丰度与对照组的腺泡和导管细胞明显不同。NKCC1 定位于所有腺泡和导管细胞的基底外侧膜，导管细胞的染色强度较弱，IAD 兔的染色模式与对照组相似。CFTR 作为点状聚集物存在于所有腺泡和导管细胞的顶细胞质中，导管细胞中的强度更强，对照组和 IAD 兔之间无显著差异。ClC2γ 也定位于所有腺泡细胞的顶细胞质作为点状聚集物，但不在导管细胞中，与对照组相比，IAD 兔中观察到类似的染色模式。
结论
我们的数据表明，IAD 兔的 NKCC1、CFTR 和 ClC2γ 的 mRNA 和蛋白表达发生了显著变化，表明这些变化可能导致 IAD 兔泪液分泌的改变，特别是 Cl 转运的改变。
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