基质金属蛋白酶-9在人正常角膜及真菌性角膜溃疡中的基因表达

Gene expression of MMP-9 in fungal corneal ulceration and normal human cornea

目的:研究基质金属蛋白酶-9(matrix metalloproteinase-9,MMP-9)在人正常角膜及真菌性角膜溃疡中的基因表达,探讨MMP-9在真菌性角膜溃疡形成中的作用。方法:搜集24份真菌性角膜溃疡标本(溃疡组)及12份人正常角膜组织标本(对照组),其中12份真菌性角膜溃疡组织和6份正常的角膜组织分别抽提组织总的RNA,以Primer5.0软件设计MMP-9的特异引物,并对标本RNA进行RT-PCR扩增,以MMP-9目的电泳条带与β-actin内参照条带灰度值比值代表相对表达量。分12份角膜溃疡组织和6份正常的角膜组织进行免疫组化检测MMP-9表达,通过免疫组化染色的灰度值进行统计学分析。结果:MMP-9在真菌性角膜溃疡中的表达明显高于正常角膜,且主要位于角膜上皮层、基底膜以及基质层。结论:真菌性角膜溃疡组织中MMP-9的基因表达明显上调。MMP-9在真菌性角膜溃疡的发生、发展及角膜穿孔中起着重要作用。加强MMP-9在真菌性角膜溃疡中的作用机制的研究,有望对真菌性角膜溃疡的治疗提供新的途径。

AIM： To study the gene expression of matrix metalloproteinase-9 （MMP-9） in fungal corneal ulceration and normal human cornea and to explore the mechanism of gene expression of MMP-9 in fungal corneal ulceration. METHODS： Twelve fungal corneal ulceration specimes and 6 normal human corneas were collected and the total RNA was extracted by Trizol veagent. Specific primers were designed using primer 5.0 software to amplify the gene fragments corresponding to MMP-9 by reverse transcription-polymerase chain reaction （RT-PCR）. The relative intensities of gene expression were estimated by the densitometry using. β-actin was as the internal control. MMP-9 was tested with immunohistochemical dying in other 12 fungal corneal ulceration specimes and 6 normal corneas. The results were analyzed by computer analysis system. RESULTS： The gene expression level of MMP-9 in fungal corneal ulceration specimes was remarkably higher than that of normal human cornea specimes. And MMP-9 mainly located in corneal epithelium and basement membrane. CONCLUSION： The expression of MMP-9 is upregulated in fungal corneal ulceration. Over transcription of MMP-9 may play an important role in pathogenesis of corneal ulceration. To emphasize the research on the mechanism of MMP-9 in provide us a new therapy the future. fungal corneal ulceration may to fungal corneal ulceration in