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正常、感染铜绿假单胞菌以及化学烧伤的小鼠角膜中半乳糖凝集素的指纹图谱分析。

Fingerprinting of galectins in normal, P. aeruginosa-infected, and chemically burned mouse corneas.

作者信息

Chen Wei-Sheng, Cao Zhiyi, Truong Laetitia, Sugaya Satoshi, Panjwani Noorjahan

机构信息

Program in Cell, Molecular & Developmental Biology, Sackler School of Graduate Biomedical Sciences, Tufts University, Boston, Massachusetts, United States.

New England Eye Center/Department of Ophthalmology, Tufts University, Boston, Massachusetts, United States.

出版信息

Invest Ophthalmol Vis Sci. 2015 Jan 6;56(1):515-25. doi: 10.1167/iovs.14-15338.

Abstract

PURPOSE

In this study, we aimed to assess whether the expression pattern of galectins is altered in Pseudomonas aeruginosa-infected and chemically burned mouse corneas.

METHODS

Galectin (Gal) fingerprinting of normal, P. aeruginosa-infected, and silver nitrate-cauterized corneas was performed by Western blotting, immunofluorescence staining, and qRT-PCR.

RESULTS

In normal corneas, Gal-1 was distributed mainly in the stroma, Gal-3 was localized mainly in epithelium, and Gal-7, -8, and -9 were detected in both corneal epithelium and stroma. Expression levels of the five galectins were drastically altered under pathological conditions. In both infected and cauterized corneas, overall Gal-3 expression was downregulated, whereas overall Gal-8 and -9 were upregulated. Changes in the expression level of Gal-7, -8, and -9 were distinct in the epithelium of infected and cauterized corneas. Expression of these three galectins was upregulated in corneal epithelium of infected corneas but not in cauterized corneas. Consistent with the changes in protein expression: (1) Gal-7, -8, and -9 mRNA expression was upregulated in cauterized corneas, and (2) Gal-3 mRNA was downregulated and Gal-9 mRNA expression was upregulated in infected corneas.

CONCLUSIONS

Our data demonstrate differential regulation of various members of the galectin family in the course of corneal infection and neovascularization. The emerging functionality of the sugar code of cell surface receptors via endogenous galectins reflect to the pertinent roles of the five tested galectins in the diseases of cornea.

摘要

目的

在本研究中,我们旨在评估在铜绿假单胞菌感染和化学灼伤的小鼠角膜中半乳糖凝集素的表达模式是否发生改变。

方法

通过蛋白质免疫印迹、免疫荧光染色和定量逆转录聚合酶链反应对正常、铜绿假单胞菌感染和硝酸银烧灼的角膜进行半乳糖凝集素(Gal)指纹图谱分析。

结果

在正常角膜中,Gal-1主要分布于基质,Gal-3主要定位于上皮,Gal-7、-8和-9在角膜上皮和基质中均有检测到。在病理条件下,这五种半乳糖凝集素的表达水平发生了显著改变。在感染和烧灼的角膜中,Gal-3的整体表达均下调,而Gal-8和-9的整体表达上调。Gal-7、-8和-9在感染和烧灼角膜上皮中的表达水平变化明显。这三种半乳糖凝集素在感染角膜的上皮中表达上调,但在烧灼角膜中未上调。与蛋白质表达变化一致:(1)Gal-7、-8和-9的mRNA表达在烧灼角膜中上调,(2)Gal-3的mRNA在感染角膜中下调,Gal-9的mRNA表达上调。

结论

我们的数据表明,在角膜感染和新生血管形成过程中,半乳糖凝集素家族的不同成员受到差异调节。通过内源性半乳糖凝集素产生的细胞表面受体糖代码的新功能反映了所检测的五种半乳糖凝集素在角膜疾病中的相关作用。

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