阿司匹林对人晶状体上皮细胞增殖及环氧合酶-2表达的影响

Effect of aspirin on cell proliferation and cyclooxygenase-2 expression in human lens epithelial cells

目的检测环氧合酶-2(cyclooxygenase-2,COX-2)在碱性成纤维细胞生长因子(basic fibroblast growth factor,bFGF)诱导的人晶状体上皮细胞(human lens epithelial cells,HLEC)增殖过程中的表达,以及阿司匹林对COX-2表达及细胞增殖的抑制作用。方法HLEC系SRA01/04传代培养,MTT比色和[3H]-TdR掺入实验检测阿司匹林对bFGF诱导的HLEC增殖和DNA合成的影响;采用逆转录聚合酶链反应和Westernblot法检测阿司匹林对COX-2mRNA和蛋白表达的影响。结果显微镜下可见,随着阿司匹林浓度增加,细胞形态学的改变逐渐明显。5mmol.L-1、10mmol.L-1、20mmol.L-1阿司匹林可明显抑制bFGF诱导的HLEC增殖,吸光度值分别是0.3662±0.0644、0.2378±0.0295、0.1898±0.0198,对照组为0.5200±0.0184,3组与对照组比较,差异均有统计学意义(均为P<0.001),抑制作用呈浓度依赖性。[3H]-TdR掺入实验结果显示,阿司匹林对晶状体上皮细胞DNA合成具有抑制作用,药物浓度与[3H]-TdR掺入量呈负相关。逆转录聚合酶链反应及Westernblot结果提示10μg.L-1bFGF组电泳COX-2mRNA及蛋白呈阳性表达,10mmol.L-1阿司匹林与10μg.L-1bFGF共同作用组电泳未见有相应条带出现。结论阿司匹林能有效抑制bFGF诱导的HLEC的生长,这种作用可能是通过抑制COX-2的表达而实现的。

Objective To investigate the expression of cyclooxygenase-2（COX-2） in the proliferation of human lens epithelial cell（HLEC） induced by basic fibroblast growth factor（bFGF）,and inhibitive effect of aspirin on the proliferation and COX-2 expression.Methods The HLEC lines SRA01/04 was serial subcultured.MTT and [^3H]-thymidine incorporation method were used to observe the effect of aspirin on the proliferation of cultured cells and the synthesis of DNA induced by bFGF.RT-PCR and Western blot were employed to detect the effect of aspirin on the expression of COX-2 mRNA and protein.Results The cellular morphous obvious changed gradually with the increased concentration of aspirin under microscope.Aspirin with the concentration of 5 mmol·L^-1,10 mmol·L^-1,20 mmol·L^-1 inhibited the proliferation of HLEC induced by bFGF obviously in a dose-dependent manner,the optical density value of them were 0.366 2±0.064 4,0.237 8±0.029 5 and 0.189 8±0.019 8 respectively,which in control group was 0.520 0±0.018 4,there were significant differences for the three groups compared with control group（all P〈0.001）.[^3H]-thymidine incorporation method showed that aspirin had the inhibitive effect on the synthesis of DNA of HLEC,the concentration of aspirin was negatively correlated with the quantity of [^3H]-thymidine incorporation.The results of RT-PCR and Western blot showed that expression of COX-2 mRNA and protein was positive in HLEC after treated with 10 μg·L^-1 bFGF,and negative after co-treated with 10 mmol·L^-1 aspirin.Conclusion Aspirin can effectively inhibit the proliferation of HLEC induced by bFGF,which is achieved by inhibiting the expression of COX-2.