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目的 探讨角膜感染Ⅰ型单纯疱疹病毒 (HSV 1)后基质金属蛋白酶 (MMPs)及其组织抑制剂 (TIMPs)在角膜中的分布及酶活性表达。方法 BALB/c小鼠眼角膜接种HSV 1(KOS株 )以诱发单纯疱疹病毒性角膜炎 (HSK)。分别收集正常眼球及感染后第 2、7、14及 2 8天的感染眼球。应用免疫组织化学法和Westernblot方法检测MMP 2、 8、 9及TIMP 1、 2在角膜组织中的表达 ,并应用酶谱(Zymography)技术检测MMPs的酶活性。结果 感染后第 2天 ,感染眼的MMP 2、 9及TIMP 1、 2表达比未感染眼增加且表达主要位于浅表基质层及上皮下的炎性细胞中。感染后第 14和 2 8天可见坏死性角膜炎及角膜溃疡形成 ,同时角膜基质和浸润的炎性细胞中尤其溃疡处 ,可见MMP 2、 9及TIMP 1、 2表达显著增加。溃疡区域有大量MMP 8阳性染色的中性粒细胞。角膜感染HSV 1后 ,明胶酶 (MMP 2、 9)活性和胶原酶 (MMP 8)活性均增强。结论 HSV 1角膜感染后 ,由角膜细胞和浸润的炎性细胞分泌产生的MMPs可能对上皮性角膜炎与溃疡形成过程起重要的促进作用。MMPs与TIMPs的相互作用可能对HSK的坏死性病变起重要调节作用。
Objective To investigate the distribution and enzyme activity of matrix metalloproteinases (MMPs) and tissue inhibitors (TIMPs) in the cornea after corneal infection with herpes simplex virus type 1 (HSV 1). Methods BALB / c mouse corneas were inoculated with HSV 1 (KOS strain) to induce herpes simplex keratitis (HSK). Eyeballs were collected at 2, 7, 14 and 28 days after infection. The expressions of MMP 2, 8, 9 and TIMP 1, 2 in corneal tissues were detected by immunohistochemistry and Western blotting. The enzymatic activities of MMPs were detected by Zymography. Results On the second day after infection, MMP 2, MMP-9 and TIMP-1, 2 in infected eyes were higher than those in non-infected eyes and mainly expressed in the superficial stromal layer and subepithelial inflammatory cells. On the 14th and 28th days after infection, necrotic keratitis and corneal ulcer were observed. At the same time, especially in the corneal stroma and infiltrating inflammatory cells, the expressions of MMP 2, MMP 9 and TIMP 1, 2 were significantly increased. There are a large number of MMP 8 positive stained neutrophils in the ulcer area. After corneal infection of HSV 1, the activities of MMP 2 and 9 and the activity of collagenase (MMP 8) increased. Conclusion The MMPs secreted by corneal cells and infiltrating inflammatory cells after HSV 1 corneal infection may play an important role in the process of epithelial keratitis and ulcer formation. The interaction between MMPs and TIMPs may play an important regulatory role in the necrosis of HSK.