Roles for Redox Signaling by NADPH Oxidase in Hyperglycemia-Induced Heme Oxygenase-1 Expression in the Diabetic Retina
Roles for Redox Signaling by NADPH Oxidase in Hyperglycemia-Induced Heme Oxygenase-1 Expression in the Diabetic Retina
复制标题
NADPH 氧化酶氧化还原信号在高血糖诱导的糖尿病视网膜血红素加氧酶-1 表达中的作用。
DOI：
10.1167/iovs.13-12004
复制
发表时间：
2013-06-01
影响因子：
4.4
通讯作者：
Pu, Mingliang
中科院分区：
文献类型：
--
作者：
He, Meihua;Pan, Hong;Pu, Mingliang
目的。抗氧化剂反应元件（IS）介导的抗氧化剂途径在维持视网膜的氧化还原状态方面具有重要作用。在DB/DB小鼠中，介导的抗氧化剂（例如血红素加氧酶-1（HO-1））的表达尚不清楚。我们评估了HO-1在DB/DB小鼠的视网膜中的表达，并研究了NADPH氧化酶的可能作用。从8-，12周和20周DB/DB或DB/M小鼠中收获新鲜视网膜。通过二乙基检测到活性氧。通过免疫组织化学和蛋白质印迹评估了HO-1，NOX2和NOX4的表达水平。体外视网膜外植体培养物用于评估NADPH氧化酶在高葡萄糖诱导的HO-1表达中的作用。与年龄匹配的对照相比，在8周DB/DB小鼠的视网膜中，HO-1的表达增加了，而20周DB/DB小鼠的表达却降低了。同样，与年龄匹配的对照相比，在8周的视网膜中，NOX4的激活在第8周时增加到基础水平。与年龄匹配的对照相比，在8-，12周和20周DB/DB小鼠的视网膜中，NOX2的激活增加。 NADPH氧化酶抑制剂Apocynin和DPI显着阻断了HO-1表达，这是由培养的视网膜外植体中高葡萄糖水平诱导的。 DB/DB小鼠视网膜中HO-1，NOX2，NOX4的表达模式，以及NADPH氧化酶抑制剂对培养的视网膜外植体中高葡萄糖水平诱导的HO-1表达的抑制作用，这表明HO-1的表达至少是由NADPH氧化酶在这种糖尿病动物模型中介导的，至少是部分介导的。
PURPOSE
The antioxidant response element (ARE)-mediated antioxidant pathway has an important role in maintaining the redox status of the retina. The expression of ARE-mediated antioxidants, such as heme oxygenase-1 (HO-1), remains unclear in the db/db mice. We evaluated the expression of HO-1 in the retinas of db/db mice and investigated a possible role for NADPH oxidase.
METHODS
Fresh retinas were harvested from 8-, 12-, and 20-week db/db or db/m mice. Reactive oxygen species were detected by dihydroethidium. The expression levels of HO-1, Nox2, and Nox4 were evaluated by immunohistochemistry and Western blotting. In vitro retina explants culture was used to assess the role of NADPH oxidase in high glucose-induced HO-1 expression.
RESULTS
The expression of HO-1 was increased in the retinas of 8-week db/db mice, while it was decreased in 20-week db/db mice compared to age-matched controls. Similarly, the activation of Nox4 was increased in the retinas at 8 weeks and returned to basal levels at 20 weeks in db/db mice compared to age-matched controls. The activation of Nox2 was increased in the retinas of 8-, 12-, and 20-week db/db mice compared to age-matched controls. The NADPH oxidase inhibitors apocynin and DPI significantly blocked the HO-1 expression that was induced by high glucose levels in cultured retina explants.
CONCLUSIONS
The expression patterns of HO-1, Nox2, Nox4 in db/db mouse retinas, and the suppressive effects of NADPH oxidase inhibitors on the expression of HO-1 induced by high glucose levels in cultured retina explants suggest that the expression of HO-1 is, at least partially, mediated by NADPH oxidase in this diabetic animal model.