Gene profiling in murine corneas challenged with Aspergillusfumigatus

[摘要] Purpose: Fungal keratitis (FK) is a blinding infection of thecorneas and accounts for a significant portion of all keratitis in theworld but little is known about the pathogenesis of FK, especially atthe level of molecular biology. This study tried to determine the genesthat are modulated in the process and thus deserve further investigationfor understanding the pathogenesis of FK.Methods: Agilent Mouse Oligo Microarray was used to compare the geneprofiles in control corneas and in corneas challenged withheat-inactivated Aspergillus fumigatus spores. Semiquantitativereverse-transcription polymerase chain reaction (RT-PCR) andimmunohistochemistry were used to confirm the changes of interestedgenes.Results: In the 18,335 genes detected by array, 61 showed anincrease of more than one-fold in RNA expression and 48 showed adecrease of over 50%. Among the regulated genes some are known to berelated to host defense such as IL3, mannose binding lectin A (MBL-A),and postaglandin D2 synthase (Psgd) while others like Dopachrometautomerase (Dct) have never been correlated to ocular or host defense.The other 33 changed genes either have unknown functions or encodehypothetical proteins. The changes in the expression of early growthresponse 4 (Egr4), Dct, Psgd, MBL-A, and hemoglobin α adult chain1 were confirmed by using RT-PCR. MBL-A expression regulation wasfurther confirmed by using immunohistochemisty in both in vitro and invivo challenged murine corneas.Conclusions: MBL-A is among the primary responding genes during theonset of fungal keratitis. Also it was found that the microarray is auseful tool in elucidating the pathogenesis of FK.

[学科分类] 生物化学/生物物理