Ding Chuanqing, Lu Michael, Huang Jianyan
Department of Cell and Neurobiology, Keck School of Medicine, University of Southern California, Los Angeles, CA 90089-9112, USA.
Mol Vis. 2011;17:2368-79. Epub 2011 Sep 1.
To test the hypothesis that expression of Na(+)/K(+)-ATPase subunits in the lacrimal glands (LGs) of rabbits with induced autoimmune dacryoadenitis (IAD) changes.
LGs were obtained from adult female rabbits with IAD and age-matched female control rabbits. The LGs were processed for laser capture microdissection (LCM), real time RT-PCR, western blot, and immunofluorescence for the detection of mRNA and proteins of the α1, α2, β1, β2, and β3 subunits of Na(+)/K(+)-ATPase.
In the rabbits with IAD, mRNA levels of α1, β1, and β3 from whole LGs were significantly lower. In samples of acini and epithelial cells from various duct segments, collected by LCM, mRNA levels of α1, β1, β2, and β3 were significantly lower in the rabbits with IAD, although mRNA for α2 could not be detected. However, western blots demonstrated that all five subunits were significantly higher in the rabbits with IAD, although their distribution patterns were similar to those of the control rabbits, as demonstrated by immunofluorescence.
The data presented herein demonstrated significant changes in mRNA and protein expressions of Na(+)/K(+)-ATPase subunits in rabbits with IAD, suggesting that these changes may play a role in the pathogenesis of Sjögren's syndrome and altered LG secretion, as observed in these animals.
验证诱导性自身免疫性泪腺炎（IAD）家兔泪腺（LG）中Na(+)/K(+)-ATP酶亚基表达发生改变这一假说。
从患有IAD的成年雌性家兔和年龄匹配的雌性对照家兔获取泪腺。对泪腺进行激光捕获显微切割（LCM）、实时逆转录聚合酶链反应（RT-PCR）、蛋白质印迹法以及免疫荧光检测，以检测Na(+)/K(+)-ATP酶α1、α2、β1、β2和β3亚基的mRNA和蛋白质。
在患有IAD的家兔中，整个泪腺的α1、β1和β3的mRNA水平显著降低。通过LCM收集的来自不同导管段的腺泡和上皮细胞样本中，患有IAD的家兔α1、β1、β2和β3的mRNA水平显著降低，尽管未检测到α2的mRNA。然而，蛋白质印迹法显示，患有IAD的家兔中所有五个亚基均显著升高，尽管免疫荧光显示它们的分布模式与对照家兔相似。
本文提供的数据表明，患有IAD的家兔中Na(+)/K(+)-ATP酶亚基的mRNA和蛋白质表达发生了显著变化，提示这些变化可能在干燥综合征的发病机制以及这些动物中观察到的泪腺分泌改变中起作用。
