Increased Expression of Cathepsins and Obesity-Induced Proinflammatory Cytokines in Lacrimal Glands of Male NOD Mouse
Increased Expression of Cathepsins and Obesity-Induced Proinflammatory Cytokines in Lacrimal Glands of Male NOD Mouse
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DOI：
10.1167/iovs.09-4523
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发表时间：
2010-10-01
影响因子：
4.4
通讯作者：
Hamm-Alvarez, Sarah F.
中科院分区：
文献类型：
--
作者：
Li, Xiaodong;Wu, Kaijin;Hamm-Alvarez, Sarah F.
目的.干燥综合征（SjS）模型雄性NOD小鼠的泪腺（LG）表现出免疫细胞浸润和脂质沉积。通过与健康对照小鼠的基因表达谱进行比较，进一步研究了SjS的机制。在蛋白水平上进一步研究差异表达的基因，以将位置和丰度的变化与疾病的发展相关联。采用基因芯片和实时荧光定量PCR技术，比较12周龄雄性NOD小鼠LG和配对BALB/c小鼠LG基因表达的差异，并采用免疫荧光和Western blot技术对目的蛋白进行定位和定量。酶法测定组织蛋白酶的催化活性。组织蛋白酶H（Ctsh）、S（Ctss）和Z（Ctsz）以及促炎因子，包括肿瘤坏死因子（Tnf）、白细胞介素6（IL 6）和白细胞介素1 β（IL 1b），在mRNA水平上调。NOD小鼠LG腺泡细胞和CD 68阳性浸润巨噬细胞的溶酶体和分泌囊泡样细胞器中检测到组织蛋白酶S免疫荧光增强。NOD小鼠LG裂解液中组织蛋白酶S（CATS）和组织蛋白酶H（CATH）活性显著高于对照裂解液，NOD小鼠泪液中CATS也显著升高。在NOD小鼠疾病模型中，CATS和CATH的表达在自身免疫性炎性疾病的发展过程中与促炎细胞因子平行增加。泪液CATS可能是诊断SjS泪腺炎的生物标志物。（Invest Ophthalmol维斯科学。2010;51：5019- 5029）DOI：10.1167/iovs.09-4523
PURPOSE. Lacrimal glands (LGs) of male NOD mice, a model of Sjogren's syndrome (SjS), exhibit immune cell infiltration and lipid deposition. The mechanism of SjS was further investigated by characterizing gene expression profiles of NOD mouse LGs in comparison with those of healthy control mice. Differentially expressed genes were further investigated at the protein level to correlate changes in location and abundance with development of disease.METHODS. Microarray followed by real-time RT-PCR was conducted to compare the gene expression in 12-week-old male NOD mouse LG relative to that in matched BALB/c mouse LG. Immunofluorescence and Western blot analyses were used to localize and quantify proteins of interest. Enzymatic assays measured catalytic activity of cathepsins.RESULTS. Cathepsin H (Ctsh), S (Ctss), and Z (Ctsz) and proinflammatory factors, including tumor necrosis factor (Tnf), interleukin 6 (Il6), and interleukin 1 beta (Il1b), were upregulated at the mRNA level. Increased cathepsin S immunofluorescence was detected in lysosomes and secretory vesicle-like organelles in LG acinar cells and CD68-positive infiltrating macrophages in NOD mouse LG. Cathepsin S (CATS) and cathepsin H (CATH) activities were significantly higher in NOD mouse LG lysate than in control lysates, and CATS was also significantly elevated in NOD mouse tears.CONCLUSIONS. Expression of CATS and CATH increases in parallel with proinflammatory cytokines during the development of autoimmune inflammatory disease in the NOD mouse disease model. Tear CATS may represent a biomarker for diagnosis of dacryoadenitis in SjS. (Invest Ophthalmol Vis Sci. 2010;51:5019- 5029) DOI:10.1167/iovs.09-4523