Pérez-Rico Consuelo, Pascual Gemma, Sotomayor Sandra, Asúnsolo Ángel, Cifuentes Alberto, García-Honduvilla Natalio, Buján Julia
Department of Surgery, Medical and Social Sciences, Faculty of Medicine and Health Sciences, University of Alcalá, Alcalá de Henares, Madrid, Spain Department of Ophthalmology, University Hospital Principe de Asturias, Alcala de Henares, Madrid, Spain.
Department of Medicine and Medical Specialities, Faculty of Medicine and Health Sciences, University of Alcalá, Alcalá de Henares, Madrid, Spain Networking Research Center on Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Madrid, Spain.
Invest Ophthalmol Vis Sci. 2014 Sep 2;55(10):6309-18. doi: 10.1167/iovs.14-14214.
We evaluated the expression of several extracellular matrix constituents implicated in the synthesis and reticulation of elastin in human pterygium, according to age and sex of the patients.
Pterygia and normal conjunctiva samples were divided into groups according to age (<50/≥50 years) and sex. Tissue was subjected to immunohistochemical staining with anti-lysyl oxidase (LOX), lyxyl oxidase-like 1 (LOXL-1), fibulin (FBLN)-5 and FBLN4, and fibrillin-1 (FBN1) antibodies. Specific primers for the same constituents were used in a quantitative real-time PCR (qRT-PCR) analysis to determine gene expression.
The LOXL-1 (P = 0.0002), FBLN5 (P = 0.0035), and FBN1 (P < 0.0001) mRNAs were significantly higher in pterygium than conjunctiva. No differences were found for LOX and FBLN4 mRNA. The expression of LOXL-1 was not affected by age or sex; however, pterygium from men and patients over 50 years old exhibited significantly higher FBLN5/FBN1 expression than did controls. The LOX gene expression was higher in the pathologic samples from the over 50-year-olds compared to the conjunctiva (P = 0.0396) and in men's versus women's pterygium (P = 0.0173). In general, the levels of LOX, LOXL-1, and FBLN5 decreased with age in healthy samples, while the pathology seemed to have increased expression of the three proteins, and even more so in older patients. The FBLN4 and FBN1 immunostaining was slight in all samples, displaying no differences between groups.
Several extracellular matrix constituents, LOXs, FBN1, and FBLN5, implicated in the development of elastin, are overexpressed in the subepithelial connective tissue extracellular matrix of human pterygium, supporting our hypothesis that elastic synthesis and reticulation is dysregulated in this type of pathology.
我们根据患者的年龄和性别,评估了几种参与人翼状胬肉中弹性蛋白合成与网状化的细胞外基质成分的表达情况。
将翼状胬肉和正常结膜样本按年龄(<50/≥50岁)和性别分组。组织用抗赖氨酰氧化酶(LOX)、赖氨酰氧化酶样1(LOXL-1)、纤维连接蛋白(FBLN)-5、FBLN4和原纤维蛋白-1(FBN1)抗体进行免疫组织化学染色。使用相同成分的特异性引物进行定量实时PCR(qRT-PCR)分析以确定基因表达。
翼状胬肉中LOXL-1(P = 0.0002)、FBLN5(P = 0.0035)和FBN1(P < 0.0001)的mRNA水平显著高于结膜。未发现LOX和FBLN4 mRNA有差异。LOXL-1的表达不受年龄或性别的影响;然而,男性和50岁以上患者的翼状胬肉中FBLN5/FBN1表达显著高于对照组。与结膜相比,50岁以上患者病理样本中的LOX基因表达更高(P = 0.0396),男性翼状胬肉中的LOX基因表达高于女性(P = 0.0173)。总体而言,健康样本中LOX、LOXL-1和FBLN5的水平随年龄下降,而在病理状态下这三种蛋白的表达似乎增加,在老年患者中更是如此。所有样本中FBLN4和FBN1免疫染色均较轻微,各分组之间无差异。
几种参与弹性蛋白发育的细胞外基质成分,即LOXs、FBN1和FBLN5,在人翼状胬肉的上皮下结缔组织细胞外基质中过表达,支持了我们的假设,即这种类型的病理状态下弹性蛋白合成和网状化失调。