Substance P in the corneal stroma regulates the severity of herpetic stromal keratitis lesions.
Substance P in the corneal stroma regulates the severity of herpetic stromal keratitis lesions.
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DOI：
10.1167/iovs.11-8089
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发表时间：
2011-11
影响因子：
4.4
通讯作者：
Brandon S. Twardy;Rudragouda Channappanavar;S. Suvas
中科院分区：
文献类型：
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作者：
Brandon S. Twardy;Rudragouda Channappanavar;S. Suvas
目的探讨单纯疱疹病毒1型（HSV-1）感染的角膜中P物质（SP）是否调节单纯疱疹性角膜基质炎（HSK）的严重程度。方法C57 BL/6小鼠眼部感染HSV-1（RE）。感染后第15天，根据角膜混浊程度分为轻度（≤2）和重度（>2）。用酶免疫吸附试验（EIA）和共聚焦显微镜检测轻、重度HSK角膜中SP的含量。在HSK临床阶段进行结膜下接种SP受体拮抗剂Spantide I。采用ELISA和流式细胞术检测角膜病变中细胞因子、趋化因子的水平和免疫细胞类型的流入。结果：与轻度角膜病变相比，重度HSK角膜病变的SP水平显著升高。与SP水平较低的角膜相比，SP水平较高的角膜也表现出较高量的促炎细胞因子（IL-6，IFN-γ）和趋化因子（CCL 3，CXCL 2）。在感染角膜的CD 45-和CD 45+细胞中测定SP受体NK 1 R的表达。SP存在于严重HSK病变眼的角膜基质中，与β-III微管蛋白（+）和IA（B+）细胞类型共定位。在HSK的临床阶段，结膜下接种Spantide I导致角膜混浊和血管生成显著减少。总之，这些结果证明了P物质在调节小鼠模型HSK病变临床严重程度中的相对贡献。
PURPOSE To determine whether substance P (SP) in herpes simplex virus-1 (HSV-1) infected cornea regulates the severity of herpetic stromal keratitis (HSK) lesions in a mouse model. METHODS C57BL/6 mice were infected ocularly with HSV-1 (RE). The corneas with HSK lesions, on Day 15 postinfection, were grouped on the basis of the corneal opacity as mild (≤2) or severe (>2). The amount of SP was determined in the corneas with mild or severe HSK lesions by enzyme immunosorbent assay (EIA) and confocal microscopy. Subconjunctival inoculation of spantide I, SP receptor antagonist, was carried out during the clinical phase of HSK. ELISA and flow cytometry were used to determine the level of cytokines, chemokines, and influx of immune cell types in the corneal lesions. RESULTS The authors determined a significantly higher level of SP in the corneas with severe HSK lesions in comparison with mild lesions. The corneas with a higher level of SP also exhibited higher amounts of proinflammatory cytokines (IL-6, IFN-γ) and chemokines (CCL3, CXCL2) when compared with the corneas with a lower level of SP. SP receptor NK1R expression was determined in CD45- and CD45+ cells in infected cornea. SP present in the corneal stroma of the eyes with severe HSK lesions colocalized with β-III tubulin(+) and IA(b+) cell types. Subconjunctival inoculation of spantide I during the clinical phase of HSK resulted in significant reduction in the corneal opacity and angiogenesis. CONCLUSIONS Collectively, these results demonstrate the relative contribution of substance P in regulating the clinical severity of HSK lesions in a mouse model.