Decreased levels of the goblet cell mucin MUC5AC in tears of patients with Sjögren syndrome.
Decreased levels of the goblet cell mucin MUC5AC in tears of patients with Sjögren syndrome.
复制标题
DOI：
--
复制
发表时间：
2002-04
影响因子：
4.4
通讯作者：
P. Argüeso;M. Balaram;M. Balaram;S. Spurr-Michaud;H. Keutmann;R. Dana;R. Dana;I. Gipson
中科院分区：
文献类型：
--
作者：
P. Argüeso;M. Balaram;M. Balaram;S. Spurr-Michaud;H. Keutmann;R. Dana;R. Dana;I. Gipson
目的确定干燥综合征患者结膜上皮中粘蛋白mRNA和泪液中粘蛋白蛋白的相对量与健康人相比是否发生了变化。方法取正常人（n = 17）和干燥综合征患者（n = 11）的下穹窿，滴入60 μ L无菌水后，收集泪液。泪液收集后，立即通过滤纸剥离从球颞区获得结膜上皮用于mRNA分离。凝胶形成粘蛋白MUC 5AC和跨膜粘蛋白MUC 1和MUC 4的非串联重复序列的引物被用于实时RT-PCR，以确定与正常受试者相比，干燥综合征患者MUC mRNA的相对丰度。酶联免疫吸附试验进行神经氨酸酶处理的眼泪，使用多克隆抗体对合成肽模仿推导的氨基酸序列从D3区MUC 5AC。结果干燥综合征患者结膜上皮中杯状细胞特异性粘蛋白MUC 5AC的RNA转录本数量显著低于正常人。当分析结膜复层上皮MUC 1和MUC 4表达的粘蛋白mRNA水平时，未检测到显着变化。杯状细胞粘蛋白MUC 5AC的蛋白水平在干燥综合征患者的泪液中显著降低，证实了使用实时RT-PCR获得的mRNA数据。结论：干燥综合征患者泪液中杯状细胞特异性粘蛋白MUC 5AC水平降低，这与结膜MUC 5AC mRNA水平降低相关。作者提出泪液中MUC 5AC粘蛋白的缺乏是导致干燥综合征泪膜不稳定的机制之一。
PURPOSE To determine whether the relative amounts of mucin mRNA in the conjunctival epithelium and mucin protein in the tears are altered in patients with Sjögren syndrome compared with healthy individuals. METHODS Tear fluid was collected from the inferior fornix of normal subjects (n = 17) and patients with Sjögren syndrome (n = 11) after instillation of 60 microL sterile water onto the ocular surface. Immediately after tear fluid collection, conjunctival epithelium was obtained by filter paper-stripping from the bulbar temporal region for mRNA isolation. Primers to nontandem repeat sequences of the gel-forming mucin MUC5AC and the membrane-spanning mucins MUC1 and MUC4 were used in real-time RT-PCR to determine relative abundance of MUC mRNA in patients with Sjögren syndrome in relation to that of normal subjects. Enzyme-linked immunosorbent assay was performed on neuraminidase-treated tears, using a polyclonal antibody against a synthetic peptide mimicking the deduced amino acid sequence from the D3 region of MUC5AC. RESULTS The number of RNA transcripts for the goblet cell-specific mucin MUC5AC in the conjunctival epithelium of patients with Sjögren syndrome was significantly lower than in normal individuals. No significant changes were detected when analyzing the mRNA levels of the mucins expressed by the stratified epithelium of the conjunctiva, MUC1 and MUC4. Protein levels of the goblet cell mucin MUC5AC were significantly reduced in the tear fluid of patients with Sjögren syndrome, corroborating mRNA data obtained using real-time RT-PCR. CONCLUSIONS The tear fluid of patients with Sjögren syndrome has reduced levels of the goblet cell-specific mucin MUC5AC, which correlates to decreased levels of conjunctival MUC5AC mRNA. The authors propose that deficiency of MUC5AC mucin in tears constitutes one of the mechanisms responsible for tear film instability in Sjögren syndrome.