【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、IκBα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 mRNA 6h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-κB p65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-kB p65的基因表达下调，核易位抑制， IκBα的基因表达上调。

【关键词】  NF-κB，IκB　前列腺素　葡萄膜巩膜

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

    Jian-Hui Xiao1, Yu-Qing Lan1, Jian-Liang Zheng2, Zhao-Xia Xia1, Chi Zhang1, Yu-Xin Hu1

    Foundation item: Doctoral Start-up of Guangdong Natural Science Fundation, China (No.04300243)

    1 Department of Ophthalmology, the Second Affiliated Hospital of Sun Yat-Sen University, Guangzhou 510120, Guangdong Province, China;2Zhongshan Ophthalmic Center, Sun Yat-Sen University, Guangzhou 510060, Guangdong Province, China

    Abstract AIM: To explore the molecular regulation of nuclear transcription factor NF-κB and its inhibitor IκB 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-κB p65 mRNA reduced in group 6 hours, group 12 hours and group 24 hours when compared with the controls（F=17.068, P=0.001）. IκBα mRNA showed no significant 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=15.4,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.

    · KEYWORDS: NF-κB; IκB; prostaglandin; uveoscleral

　　0引言

    葡萄膜巩膜房水外流通道是房水排出的重要途径之一，许多研究表明，前列腺素类抗青光眼药物是通过增加葡萄膜巩膜房水外流从而降眼压。睫状肌在葡萄膜巩膜房水流出通道中有关键作用，是葡萄膜巩膜房水外流的限速部位[1]。核转录因子NF-кB（nuclear factor kappa B）是近年发现的一种重要的转录因子。NF-кB系统由NF-кB家族及其抑制物IкB家族共同组成[2,3]。NF-кB家族由Rel蛋白家族中的成员以同源或异源二聚体的形式存在。NF-кB存在于多种细胞中，参与炎症、免疫、应激反应、细胞增殖和细胞凋亡等多种生理、病理过程的基因调控[4,5]。研究表明，NF-кB作为广泛存在于各种细胞的核转录调控因子，可能与葡萄膜巩膜房水流出通道的多种细胞信号调控有关。Travoprost是由爱尔康（中国）眼科产品有限公司研制的一种新型前列腺素类抗青光眼药物。其作用下人睫状肌细胞中NF-кB的变化目前尚未见报道。

[1] [2] [3] [4] 下一页