曲伏前列腺素对培养人眼睫状肌细胞基质金属蛋白酶-2表达的影响  

Effects of Travoprost on Matrix Metalloproteinses-2 in Cultured Human Ciliary Muscle Cells

作　　者：蓝育青[1] 肖剑晖[1] 彭蔚[1] 张弛[1] 郭慧[1] 宋欣[1] 

机构地区：[1]中山大学附属第二医院眼科,广东广州510120

出　　处：《中山大学学报（医学科学版）》2007年第5期498-500,514,共4页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：广东省自然科学基金博士启动基金(04300243)

摘　　要：【目的】研究曲伏前列腺素作用下,基质金属蛋白酶-2(MMP-2)在人睫状肌细胞的表达。【方法】在人睫状肌细胞无牛血清培养基中加入1μmol/L曲伏前列腺素(travoprost),根据曲伏前列腺素孵育时间的不同,分为4个时间组,即0 h组(对照组)、6 h、12 h、24 h组。Real-time PCR和ELISA法分别检测上述各时间组人睫状肌细胞MMP-2在基因和蛋白水平的表达,每种方法分别重复做3次。Zymography技术分别检测4组细胞MMP-2的活性,重复4次。【结果】设定对照组mRNA相对表达量为1做作标准,6 h、12 h、24 h组MMP-2 mRNA相对表达量为0.58±0.04、1.20±0.05、1.95±0.11,MMP-2 mRNA表达呈逐渐升高趋势(P=0.000);ELISA检测MMP-2的A值分别为0.0503±0.0021、0.0627±0.0017、0.0673±0.0025、0.0783±0.0039,MMP-2的表达随travoprost作用时间延长逐渐升高(P=0.000)。Zymography技术检测MMP-2校正光密度值分别为10±5,52±7,104±21,237±40;MMP-2活性随travoprost作用时间延长而逐渐增强(P=0.000)。【结论】曲伏前列腺素作用于人睫状肌细胞后,MMP-2表达随药物作用时间延长逐渐增加,活性逐渐增强。[Objective ]To study the expression of matrix metalloproteinase-2 （MMP-2） in human ciliary muscle cells after administration of travoprost. [ Methods ] 1 μmol/L travoprost was diluted in the serum-free cultured medium of human ciliary muscle cells （HCM）. Four groups which including group 0 h （control group）, group 6 h, group 12 h, and group 24 h were determined according to the exposed time. Real-time PCR and ELISA were used to detect the relative quantitative mRNA and expression of protein of MMP-2 in the 4 groups, respectively. Each method was repeated for 3 times. Zymography was used to detect the activity of MMP-2 in the HCM cells of 4 groups for 4 times. [ Results ] When considering the result of relative quantitative mRNA of control group as 1, the results of relative quantitative mRNA of 6 h, 12 h, and 24 h were 0.58±0.04, 1.20±0.05, and 1.95±0.11, respectively. Expression of MMP-2 in mRNA level was showed a gradually increasing trend （P =0.000）. The value A of 4 groups with ELISA were 0.0503±0.0021, 0.0627±0.0017, 0.0673±0.0025, and 0.0783±0.0039, respectively, which revealed that the expression of MMP-2 in protein level increased gradually with the exposed time （P=0.000）. With zymographic analysis, the corrected ray density of 4 groups were 10±5, 52±7, 104±21, and 237±40, respectively. The activity of MMP-2 improved gradually with the exposed time （P=0.000）. [Conclusion] The expression of MMP-2 in mRNA and protein level increased gradually and the activity of MMP-2 improved with the exposed time of travoprost in human ciliary muscle cells.

关 键 词：睫状肌细胞 基质金属蛋白酶 曲伏前列腺素 基因表达 

分 类 号：R77[医药卫生—眼科]