RESUMO
This study aimed to investigate expressions and sources of matrix metalloproteinases (MMP)-2 and MMP-9, and of tissue inhibitors of MMP (TIMP)-1 and TIMP-2 in experimental Pseudomonas aeruginosa keratitis in rabbits. Pseudomonal keratitis was induced in New Zealand white rabbits, and macroscopic and microscopic examinations were performed at appropriate time points (3, 9, 12, 18, 24, 72 h). Expressions and sources of MMP-2, 9, and TIMP-1, 2 were determined using immunohistochemistry, gelatin zymography, ELISA, and RT-PCR. A typical corneal ulcer with a ring abscess was observed 12-72 h post-infection (p.i.) with P. aeruginosa. In microscopic examinations, massive inflammatory cell (mostly polymorphonuclear leukocytes, PMNs) infiltration and liquefactive necrosis were characteristic features. MMP-2 was constitutively expressed in keratocytes, and its expression was not apparently enhanced after pseudomonal infection as evidenced by zymography, immunostaining, and RT-PCR. However, MMP-9 and its activated form were induced, and were significantly enhanced 12-24 h p.i. MMP-9 appeared to derive from PMNs rather than from resident corneal cells. TIMP-1 was expressed in PMNs, macrophages, and keratocytes, and its expression was enhanced 72 h p.i. Although TIMP-2 was constitutively expressed as seen by immunostaining and RT-PCR, its concentration was below detection limits during the experiments. We demonstrated that MMP-9 was one of the important factors for corneal tissue destruction, because it was induced and significantly expressed in keratocytes and inflammatory cells after pseudomonal infection. Although TIMP-1 was expressed in later stages of infection, enhancement and activation of MMP-9 were much faster and stronger than those of TIMP-1, thereby facilitating tissue destruction leading to corneal ulceration.
Assuntos
Ceratite/metabolismo , Metaloproteinases da Matriz/metabolismo , Infecções por Pseudomonas/metabolismo , Inibidores Teciduais de Metaloproteinases/metabolismo , Doença Aguda , Animais , Córnea/enzimologia , Feminino , Expressão Gênica , Ceratite/microbiologia , Ceratite/patologia , Metaloproteinase 2 da Matriz/genética , Metaloproteinase 2 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/genética , Metaloproteinase 9 da Matriz/metabolismo , Metaloproteinases da Matriz/genética , Infecções por Pseudomonas/enzimologia , RNA Mensageiro/genética , Coelhos , Inibidor Tecidual de Metaloproteinase-1/genética , Inibidor Tecidual de Metaloproteinase-1/metabolismo , Inibidor Tecidual de Metaloproteinase-2/genética , Inibidor Tecidual de Metaloproteinase-2/metabolismo , Inibidores Teciduais de Metaloproteinases/genéticaRESUMO
Pseudomonal keratitis usually progresses rapidly, often resulting in corneal perforation and blindness. Remarkable events in pseudomonal keratitis include massive polymorphonuclear leukocyte infiltration in the cornea and various degrees of tissue destruction. With regard to initiation of these inflammatory events, various inflammatory cytokines and chemokines appear to be key substances and have been the subject of several studies. Inflammatory cytokines and chemokines believed to be important in pseudomonal keratitis include interleukin (IL)-1 beta, IL-6, macrophage inflammatory protein (MIP)-2 (homologous to human IL-8), macrophage inhibitory factor (MIF), IL-12, IL-18, interferon (IFN)-gamma, and tumor necrosis factor (TNF)-alpha. In this article, current concepts related to the role of inflammatory cytokines and chemokines in pseudomonal keratitis are reviewed.
