氢气和 N-乙酰-L-半胱氨酸可挽救小鼠角膜碱烧伤模型中的氧化应激诱导的血管生成。

Hydrogen and N-acetyl-L-cysteine rescue oxidative stress-induced angiogenesis in a mouse corneal alkali-burn model.

机构信息

Department of Ophthalmology, Keio University School of Medicine, Shinjuku-ku, Tokyo, Japan.

出版信息

Invest Ophthalmol Vis Sci. 2011 Jan 21;52(1):427-33. doi: 10.1167/iovs.10-6167.

DOI:10.1167/iovs.10-6167

PMID:20847117

Abstract

PURPOSE

To investigate the role of reactive oxygen species (ROS) as the prime initiators of the angiogenic response after alkali injury of the cornea and observe the effects of antioxidants in preventing angiogenesis.

METHODS

The corneal epithelia of SOD-1-deficient mice or wild-type (WT) mice were removed after application of 0.15 N NaOH to establish the animal model of alkali burn. ROS production was semiquantitatively measured by dihydroethidium (DHE) fluorescence. Angiogenesis was visualized by CD31 immunohistochemistry. The effects of the specific NF-κB inhibitor DHMEQ, the antioxidant N-acetyl-L-cysteine (NAC), and hydrogen (H2) solution were observed.

RESULTS

ROS production in the cornea was enhanced immediately after alkali injury, as shown by increased DHE fluorescence (P<0.01). NF-κB activation and the upregulation of vascular endothelial growth factor (VEGF) and monocyte chemoattractant protein-1 (MCP-1) were significantly enhanced (P<0.01), leading to a significantly larger area of angiogenesis. Angiogenesis in SOD-1-/- mice corneas were significantly higher in WT mice (P<0.01), confirming the role of ROS. Pretreatment with the specific NF-κB inhibitor DHMEQ or the antioxidant NAC significantly reduced corneal angiogenesis by downregulating the NF-κB pathway (P<0.01) in both WT and SOD-1-/- mice. Furthermore, we showed that irrigation of the cornea with hydrogen (H2) solution significantly reduced angiogenesis after alkali-burn injury (P<0.01).

CONCLUSIONS

Immediate antioxidant therapy with H2-enriched irrigation solution is a new potent treatment of angiogenesis in cornea to prevent blindness caused by alkali burn.

摘要

目的

研究活性氧（ROS）作为角膜碱烧伤后血管生成反应的最初启动子的作用，并观察抗氧化剂在预防血管生成中的作用。

方法

用 0.15N NaOH 涂抹 SOD-1 缺陷型和野生型（WT）小鼠角膜上皮，建立碱烧伤动物模型。通过二氢乙啶（DHE）荧光半定量测量 ROS 的产生。用 CD31 免疫组化法观察血管生成。观察 NF-κB 特异性抑制剂 DHMEQ、抗氧化剂 N-乙酰-L-半胱氨酸（NAC）和氢气（H2）溶液的作用。

结果

碱损伤后角膜中 ROS 生成增加，DHE 荧光增强（P<0.01）。NF-κB 激活和血管内皮生长因子（VEGF）和单核细胞趋化蛋白-1（MCP-1）的上调明显增强（P<0.01），导致血管生成面积明显增大。SOD-1-/-小鼠角膜的血管生成明显高于 WT 小鼠（P<0.01），证实了 ROS 的作用。NF-κB 特异性抑制剂 DHMEQ 或抗氧化剂 NAC 的预处理通过下调 NF-κB 通路（P<0.01），明显降低了 WT 和 SOD-1-/-小鼠的角膜血管生成。此外，我们表明，用富含氢气（H2）溶液冲洗角膜可显著减少碱烧伤后的血管生成（P<0.01）。