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山东大学学报(医学版) ›› 2014, Vol. 52 ›› Issue (10): 40-44.doi: 10.6040/j.issn.1671-7554.0.2014.306

• 基础医学 • 上一篇    下一篇

色素上皮衍生因子对高糖状态大鼠视网膜Müller细胞NF-κB表达的影响

毕欢丽, 李艳, 张杰, 李梦云   

  1. 潍坊医学院眼科教研室, 山东 潍坊 261031
  • 收稿日期:2014-05-12 修回日期:2014-09-19 出版日期:2014-10-10 发布日期:2014-10-10
  • 通讯作者: 李艳。E-mail:liyanmails@126.com E-mail:liyanmails@126.com
  • 基金资助:
    潍坊市卫生局科研基金(2014034)

Effect of PEDF on the expression of NF-κB on high glucose-stimulated rat retinal Müller cells

BI Huanli, LI Yan, ZHANG Jie, LI Mengyun   

  1. Department of Ophthalmology, Weifang Medical College, Weifang 261031, Shandong, China
  • Received:2014-05-12 Revised:2014-09-19 Online:2014-10-10 Published:2014-10-10

摘要: 目的 探讨色素上皮衍生因子(PEDF)对高浓度葡萄糖作用下体外培养大鼠视网膜Müller细胞损伤的保护作用及对核因子-κB(NF-κB)表达的影响。方法 采用出生3~7 d的SD大鼠视网膜组织,应用酶消化法培养Müller细胞,应用免疫细胞荧光双标方法鉴定细胞。细胞随机分为正常对照组(对照组),高糖模型组(HG组)和PEDF干预高糖组(HG+PEDF组)。采用Western boltting和免疫细胞化学染色法检测NF-κB 的表达。结果 Western boltting结果可见,与对照组相比,HG组的NF-κB蛋白表达明显增强(P<0.01);与HG组相比,HG+PEDF组NF-κB蛋白表达显著降低(P<0.001)。免疫细胞化学染色法结果可见,NF-κB在对照组仅有极少量在胞浆中表达,在HG组胞浆和胞核中皆呈强阳性表达,两者差异具有统计学意义(P<0.001);而HG+PEDF组的NF-κB表达低于其在HG组中的表达(P<0.01)。结论 体外25 mmol·;L-1高糖刺激可增加大鼠视网膜Müller细胞NF-κB的表达,从而加重Müller细胞损伤;外源给予PEDF可以显著降低NF-κB的表达从而减少Müller细胞损伤,对视网膜Müller细胞具有显著的保护作用。

关键词: 色素上皮衍生因子, 核因子-κB, Mü, ller细胞, 大鼠, 高糖刺激

Abstract: Objective To investigate the protective effect of pigment epithelium-derived factor(PEDF)on cultured rat Müller cell under high concentrations of glucose, and its effect on nuclear factor-kappa B (NF-κB) expression. Methods Müller cells were cultured with retinal tissues of SD rats postnatal 3 to 7 days by enzyme digestion. The cells were identified by double-labeled immunofluorescence staining. The cells were randomly divided into normal control group (control group), high glucose model group (HG group) and PEDF-intervention high glucose group (HG+PEDF group). Western boltting method and immunocytochemical staining were used to detect the expression of NF-κB. Results Western boltting method showed the expression of NF-κB in HG group increased significantly compared with control group (P<0.01). NF-κB protein expression in HG+PEDF group decreased obviously compared with HG group(P<0.001). Immunocytochemical staining showed that only a few expressions of NF-κB in control group, while highly strong expression in cytoplasm and nucleus in HG group (P<0.001). Compared with HG group, NF-κB protein expression in HG+PEDF group decreased significantly (P<0.01). Conclusion 25 mmol·L-1 high glucose increases the expression of NF-κB in vitro, and lead to the injury of Müller cells. Exogenous PEDF can decrease the expression of NF-κB on retinal Müller cells and decrease cells' injury significantly. So PEDF has a significant protective effect on retinal Müller cells.

Key words: Pigment epithelium-derived factor, Mü, ller cells, Rat, High glucose-stimulatioin, Nuclear factor-kappa B

中图分类号: 

  • R774.1
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