【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。更多还原

【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. ·更多还原