衰老对大鼠视网膜色素上皮细胞ROS生成和NADPH氧化酶表达的影响

目的衰老对大鼠视网膜色素上皮细胞(RPE)活性氧物质(ROS)生成及对尼克酰胺腺嘌呤二核苷酸磷酸(NADPH)氧化酶表达的影响。方法实验分为青年组(3月龄)和老年组(16月龄)大鼠,二氢乙啡啶(DHE)染色检测RPE ROS的生成,HE染色观察RPE形态学变化,免疫组化检测视网膜及RPE蛋白激酶Cα(PKCα)、P47phox和NADPH氧化酶2(NOX2)的表达。结果与青年组大鼠比较,老年组大鼠RPE ROS的生成明显增加。HE染色结果显示,青年组和老年组大鼠视网膜及RPE形态学未见明显差异。免疫组化结果表明,老年组大鼠视网膜及RPE PKCα、P47phox和NOX2的表达明显增加。结论老年大鼠RPEROS生成和NADPH氧化酶表达明显增加,NADPH氧化酶介导的ROS可能是年龄相关性视网膜病变的机制之一。

Study of the ROS generation and expression of NADPH oxidase in the retina pigment epithelial cell in aged rats

Objective To study the reactive oxygen species(ROS) generation and expression of NADPH oxidase in the retina and pigment epithelial cells(RPE) in aged rats.Methods Rats were divided into two groups: young group(3-month old) and old group(16-month old).DHE staining was used to detect the ROS generation in pigment epithelial cells.The histological changes were investigated by HE staining.The expression of PKCα,P47phox and NOX2 in the RPE were detected by immunohistochemistry assay.Results Compared with the young group,the ROS production in pigment epithelial cells was increased significantly in the old group rats.Histology results showed that there was no significant difference in the RPE between the young and old group rats.Immunohistochemistry assay showed that,compared with young group,the expression of PKCα,P47phox and NOX2 in the RPE were significantly increased.Conclusion The ROS generation and expression of NADPH oxidase in the RPE are significantly increased in aged rats,and the ROS mediated by NADPH oxidase may be one of the mechanisms of age associated retinopathy.