NF-κB对体外培养的人睫状肌细胞的作用  

Effect of NF-κB on human ciliary muscle cells in vitro

作　　者：肖剑晖[1] 蓝育青[1] 郑健樑[2] 夏朝霞[1] 张弛[1] 胡玉新[1] 

机构地区：[1]中山大学附属第二医院眼科,中国广东省广州市510120 [2]中山大学中山眼科中心,中国广东省广州市510060

出　　处：《国际眼科杂志》2007年第1期80-82,共3页International Eye Science

基　　金：2004年中国广东省自然科学基金博士启动基金资助(No.04300243)~~

摘　　要：目的:观察PGF2α类曲伏前列腺素药物travoprost作用下,核转录因子NF-κB及其抑制因子IκB对体外培养的人睫状肌细胞的分子调控。方法:在人睫状肌细胞培养基中加1μmol/L曲伏前列腺素travoprost,根据孵育时间的不同分为4组,即(0对照组),6,12,24h组。采用real-time RT-PCR、免疫荧光半定量分析和ELISA法分别检测上述时间组NF-κB p65、Iκ-Bα在基因和蛋白水平的表达。结果:与对照组比较,6,12,24h组NF-κB p65mRNA表达均下降(F=17.068,P=0.001);IκBα mRNA6h组、12h组较对照组改变不明显(P>0.05),24h组较对照组表达增加(F=32.742,P=0.000)。免疫荧光半定量分析表明:NF-κBp65荧光强度6,12,24h组均较对照组减少(F=17.216,P=0.000);IκBα6h组较对照组没有明显改变(P=0.134)、12h组较对照组轻微下降(P=0.032),24h组较对照组明显增加(F=17.346,P=0.001);ELISA法检测磷酸化NF-κBp65随药物作用时间延长而逐渐下降(F=15.4,P=0.001)。结论:曲伏前列腺素作用于人睫状肌细胞后,NF-kBp65的基因表达下调,核易位抑制,IκBα的基因表达上调。AIM： To explore the molecular regulation of nuclear transcription factor NF-κB and its inhibitor IKB in human ciliary muscle （HCM） cells in vitro by PGF2α prostaglandin analogue travoprost. METHODS： Cultured HCM cells were divided into four groups： group 6 hours, group 12 hours, group 24 hours that were determined by the time of HCM cells being exposed to 1μmol/L travoprost and control group （travoprost free）. RT-PCR, immunofluorescence relative quantitative analyses and ELISA were used to detect the expression of NF-κB and IκBα in the four groups. RESULTS： NF-KB p65 mRNA reduced in group 6 hours, group 12 hours and group 24 hours when compared with the centrols（F=17.068, P=-0.001）. IκBα mRNA showed no significent changes in group 6h and group 12 hours （P 〉0.05）, while group 24h increased significantly compared with the controls （F=32.742, P=-0.000）. relative quantitative Immunofluorescence analyses showed that the fluorescence intensity of NF-κB p65 in group 6 hours, 12 hours and 24 hours decreased when compared with that in controls （F=17.216, P=-0.000）. It showed no significant change of fluorescence intensity in group 6h （P=0.134）, slight reduction in group 12 hours（P=0. 032） and marked increase in group 24 hours （F= 17.346,P=0.001） when compared with the control group. ELISA revealed that phosphorylation of NF-κB p65 decreased gradually with the time of exposure to travoprost in HCM （F=15A, P=0.001）. CONCLUSION： Down-regulation of NF-κB p65 mRNA and inhibition of nuclear translocation were detected while expression of IκBα increases when HCM cells are exposed to travoprost.

关 键 词：NF—κB IκBα 前列腺素 葡萄膜巩膜 

分 类 号：R773.3[医药卫生—眼科]