摘要
目的 研究肾上腺髓质素 ( adrenomedullin,ADM)对人视网膜色素上皮 ( retinal pigment epi-thelium,RPE)细胞表达核转录因子 -κB ( nuclear transcription factor-κB,NF-κB)的影响。 方法 体外培养人 RPE细胞 ,分别加入终浓度为 10 - 7mol/ L的 ADM或终浓度为 10μg/ ml的脂多糖( lipopolysaccharide,L PS) ,采用免疫荧光抗体染色、Western blotting分析 ,观察 NF-κB的蛋白表达水平。 结果 未受刺激的 RPE细胞细胞浆内呈现弱的绿色荧光。ADM或 L PS刺激 6 0 min后细胞核及核周围细胞浆可见到明亮的绿色荧光 ,染色呈强阳性。Western blotting分析结果表明未受刺激的 RPE细胞在相对分子质量为 36× 10 3处可见一条阳性带 ,表达微量的 NF- κB。加入 L PS的 RPE细胞中 NF- κB的蛋白表达量明显增多 ,加入 ADM的 RPE细胞也可明显上调 NF- κB的表达量。 结论 ADM可激活 RPE细胞内的 NF- κB,使其发生核内转移并上调细胞浆内 NF- κB的蛋白表达量 ,这可能是 ADM调控 RPE细胞的一个重要信号转导途径。
Objective To investigate the effects of adrenomedullin (ADM) on the expression of nuclear transcription factor-κB (NF-κB) in cultured human retinal pigment epithelial (RPE) cells. Methods Cultured human RPE cells were exposed to ADM with the concentration of 10 -7mol/L or lipopolysaccharide (LPS) with the concentration of 10 μg/ml for 60 minutes. Nuclear translocation of NF-κB was determined by immunofluorescence staining. Western blotting analysis was used to check the expression of NF-κB protein. Results In cultured human RPE cells that were not stimulated, weak NF-κB positive green flurescence was localized in the cytoplasm. After exposed to ADM or LPS for 60 minutes, there was bright NF-κB nuclear staining in cultured human RPE cells. The results of Western blotting analysis demonstrated that in cultured human RPE cells that were not stimulated, a single weak protein strip with the relative molecular weight of 36×103 identified as immunoreative NF-κB could be observed. Both ADM and LPS could markedly increase the expression of NF-κB protein in cytoplasm. Conclusions NF-κB-dependent signal transduction pathway can be activated in cultured human RPE cells by ADM. ADM could induce NF-κB nuclear translocation and increase the expression of the protein in cytoplasm. NF-κB-dependent signal transduction pathway may play an importantrole in the function of ADM.
出处
《中华眼底病杂志》
CAS
CSCD
2003年第3期156-159,共4页
Chinese Journal of Ocular Fundus Diseases
基金
国家自然科学基金资助项目 (39970 780 )
