脂多糖诱导人角膜基质细胞核因子-κB的活化表达及意义  

Activation and significance of NF-κB induced by lipopolysaccharide in cultured human corneal fibroblasts

作　　者：陈国玲[1,2] 陈双 刘艳丽[1] 巍璐婉[1] 刘执玉[1] 

机构地区：[1]山东大学医学院解剖学教研室,济南250012 [2]山东大学眼科中心第二医院眼科,济南250033 [3]枣庄市山亭区人民医院,山东枣庄277200

出　　处：《山东大学学报（医学版）》2009年第3期8-11,共4页Journal of Shandong University:Health Sciences

基　　金：山东省教育厅科技计划项目资助课题(J08LH59)

摘　　要：目的研究脂多糖(LPS)诱导人角膜基质细胞核因子-κBp65(NF-κBp65)和NF-κB抑制因子-α(IκB-α)的表达及意义。方法原代培养人眼角膜基质细胞,用免疫细胞化学SABC方法进行细胞鉴定。采用绿脓杆菌LPS刺激角膜基质细胞,分别于刺激前(0 h),刺激后1、2、4、8 h收集细胞。Western blot检测胞浆内IκB-α蛋白及胞核内NF-κBp65蛋白的表达。结果与0 h相比,LPS刺激细胞1 h时,即可检测到胞核内NF-κBp65蛋白表达增加,其电泳条带的光密度值随着刺激时间的延长而逐渐变亮,4 h表达最高。LPS刺激细胞后各时点的NF-κBp65蛋白表达与0 h组比较,差异有统计学意义(P均<0.01)。LPS刺激角膜基质细胞后,在各时间点均可引起胞浆IκB-α表达下降,4 h时表达最低(P均<0.01)。结论LPS可引起人眼角膜基质细胞IκB-α的降解,促进NF-κB的核转位,在角膜炎的过程中可能起一定的抗炎作用。Objective To investigate expressions and significance of nuclear factor-kappaB （NF-κB） and inhibitor of NF-κB （IκB）-αinduced by hpopolysaccharide（LPS） in cultured human corneal fibroblasts. Methods Human corneal fibroblasts were primarily cultured and identified by the SABC method. Immunocytochemical staining with an antibody against Vimentin was performed to identify these cells. Cells were collected before being challenged with LPS and 1,2,4 and 8h after LPS treated, then protein expressions of NF-κBp65 and IκB-α were assessed by Western Blot analysis. Results Compared with the 0 h group, NF- κBp65 level was significandy enhanced in the nucleus of human corneal fibroblasts after LPS challenge of 1 to 8 h（ P 〈 0.01 ）, the peak expression of NF-κBp65 was observed at 4 h. However, the protein of the IκBα level was significantly decreased after LPS-treatment at each point （ P 〈 0.01 ）. Conclusion These results suggest that LPS takes part in the degeneration of IκBα and mediates nuclear translocation of NF-κB in cultured human corneal fibroblasts in vitro.

关 键 词：脂多糖 角膜基质 NF-ΚB NF-κB抑制因子-α 

分 类 号：R772.2[医药卫生—眼科]