Murine ocular heparanase expression before and during infection with Pseudomonas aeruginosa
Murine ocular heparanase expression before and during infection with Pseudomonas aeruginosa
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DOI：
10.1167/iovs.03-0589
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发表时间：
2004-04-01
影响因子：
4.4
通讯作者：
Vlodavsky, I
中科院分区：
文献类型：
--
作者：
Berk, RS;Dong, Z;Vlodavsky, I
目的.目的研究乙酰肝素酶（硫酸乙酰肝素内切糖苷酶）在正常小鼠眼和角膜内感染铜绿假单胞菌的小鼠中的组成性表达和调控。用铜绿假单胞菌感染C57 BL/6 J小鼠，用半定量RT-PCR和免疫印迹法检测其角膜乙酰肝素酶基因和蛋白的表达。还应用免疫组织化学方法对幼稚小鼠角膜乙酰肝素酶进行了表征。结果。C57 BL/6 J小鼠未感染角膜中检测到乙酰肝素酶mRNA和蛋白表达。免疫组化结果显示，肝素酶蛋白在感染前主要表达于角膜上皮，感染后也表达于角膜基质。免疫组织化学研究未感染和感染的幼稚小鼠的整个眼睛表明乙酰肝素酶蛋白的表达，在大多数层的视网膜，但表达没有出现在角膜感染上调。视网膜内感光细胞层染色最强。在角膜内感染铜绿假单胞菌之前，乙酰肝素酶在角膜上皮和几个视网膜层中组成性表达。在感染期间，在未处理小鼠中检测到角膜乙酰肝素酶蛋白表达的时间上调，最可能是由于乙酰肝素酶阳性浸润细胞，但在免疫小鼠的角膜中蛋白质未上调，因为它们具有较低的炎症反应，与角膜透明度的恢复相关。通过免疫组织化学测定，感染小鼠视网膜中的乙酰肝素酶表达似乎没有时间上调。
PURPOSE. To demonstrate the constitutive expression and regulation of heparanase (heparan sulfate endoglycosidase) in the normal mouse eye and in mice intracorneally infected with Pseudomonas aeruginosa.METHODS. Naive (unimmunized) and immunized C57BL/6J mice were infected with P. aeruginosa, and corneal heparanase gene and protein expression were detected by semiquantitative RT-PCR and immunoblot analysis. Immunohistochemistry was also applied to characterize corneal heparanase in naive mice.RESULTS. Heparanase mRNA and protein expression were detected in uninfected corneas of C57BL/6J mice. Immunohistochemical studies indicated heparanase protein expression was primarily in the corneal epithelium before corneal infection and was also in the corneal stroma after infection. Immunohistochemical studies of uninfected and infected whole eyes of naive mice indicated heparanase protein expression in most layers of the retina, but the expression did not appear to be upregulated during corneal infection. Staining was most intense in the inner photoreceptor layer of the retina.CONCLUSIONS. Heparanase was constitutively expressed in both the corneal epithelium and several retinal layers before intracorneal infection with P. aeruginosa. Temporal upregulation of corneal heparanase protein expression was detected in naive mice during infection, most likely due to heparanase positive infiltrating cells, but the protein was not upregulated in corneas from immunized mice because they had a lower inflammatory response, associated with the restoration of corneal clarity. There did not appear to be temporal upregulation of heparanase expression in the retina of infected mice, as determined by immunohistochemistry.