Corticosteroid and doxycycline suppress MMP-9 and inflammatory cytokine expression, MAPK activation in the corneal epithelium in experimental dry eye
Corticosteroid and doxycycline suppress MMP-9 and inflammatory cytokine expression, MAPK activation in the corneal epithelium in experimental dry eye
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DOI：
10.1016/j.exer.2006.02.004
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发表时间：
2006-09-01
影响因子：
3.4
通讯作者：
Pfiugfelder, Stephen C.
中科院分区：
文献类型：
--
作者：
De Paiva, Cintia S.;Corrales, Rosa M.;Pfiugfelder, Stephen C.
探讨糖皮质激素和强力霉素对实验性干眼小鼠基质金属蛋白酶-9和炎性细胞因子的表达及丝裂原活化蛋白激酶信号转导通路c-jun氨基末端激酶、细胞外调节蛋白激酶和p38的影响。用C57BL6小鼠建立实验性干眼模型，分别给予1%甲基强的松龙、0.025%多西环素或平衡盐溶液，每日4次。激光扫描共聚焦显微镜检测角膜上皮细胞中基质金属蛋白酶-9的表达。用原位酶谱法检测角膜明胶酶活性，用明胶酶谱法检测泪液中基质金属蛋白酶-9的活性。Luminex免疫珠法检测总MAPKs和磷酸化MAPKs(JNK1/2、ERK1/2、p38)。采用实时荧光定量聚合酶链式反应(Real-Time-PCR)检测细胞内基质金属蛋白酶-9(MMP9)、白介素1α(IL-1α)、白介素1β(IL-1β)和肿瘤坏死因子(TNF)-αRNA转录水平。正常对照组角膜基质金属蛋白酶-9免疫反应定位于角膜根尖上皮细胞膜。干燥应激显著增加角膜上皮细胞产生的基质金属蛋白酶-9，并增加其在角膜上皮细胞和泪液中的活性。干燥还可增加角膜上皮细胞IL-1α、IL-1β和TNF-αmRNA的表达，刺激JNK1/2、ERK1/2和p38MAPKs的磷酸化。甲基强的松龙和强力霉素均可降低角膜上皮细胞基质金属蛋白酶-9的表达和活性，降低炎症细胞因子转录水平，减少MAPKs的激活。干燥应激刺激角膜上皮细胞表达基质金属蛋白酶-9、IL-1α、IL-1β和肿瘤坏死因子-α，并激活MAPK信号通路。皮质类固醇和多西环素都能抑制这种分子应激反应。(C)2006爱思唯尔有限公司。保留所有权利。
We investigated the effects of corticosteroid and doxycycline on expression of matrix metalloproteinase (MMP)-9 and inflammatory cytokines and activation of mitogen-activated protein kinase (MAPK) signaling pathways, c-jun N-terminal kinases (JNK), extracellular-regulated kinases (ERK) and p38, in experimental murine dry eye. Experimental dry eye (EDE) was created in C57BL6 mice, with or without or topical treatment consisting of 1% methylprednisolone, 0.025% doxycycline or balanced salt solution four times per day. MMP-9 expression in the cornea epithelia was evaluated by laser scanning confocal microscopy. Gelatinase activity in the cornea was evaluated by in situ zymography and MMP-9 activity in tear washings was evaluated by gelatin zymography. Total and phosphorylated MAPKs (JNK1/2, ERK1/2, p38) were detected by Luminex immunobead assay. Levels of MMP-9, interleukin (IL)-1 alpha, IL-1 beta and tumor necrosis factor (TNF)-alpha RNA transcripts were evaluated by real-time PCR. MMP-9 immunoreactivity was localized to the apical corneal epithelial cell membranes in normal control eyes. Desiccating stress significantly increased production of MMP-9 by the corneal epithelium and increased its activity in the corneal epithelium and tear fluid. Dryness also increased expression of IL-1 alpha, IL-1 beta and TNF-alpha mRNA and stimulated phosphorylation of JNK1/2, ERK1/2 and p38 MAPKs in the corneal epithelium. Both methylprednisolone and doxycycline reduced expression and activity of MMP-9, decreased levels of inflammatory cytokines transcripts and reduced activation of MAPKs in the corneal epithelium in response to EDE. Desiccating stress stimulates expression of MMP-9, IL-1 alpha, IL-1 beta and TNF-alpha mRNA, as well as activates MAPK signaling pathways in the corneal epithelium. Both corticosteroid and doxycycline suppressed this molecular stress response. (c) 2006 Elsevier Ltd. All rights reserved.