蓝光照射对人视网膜色素上皮细胞L-型钙通道mRNA表达的影响  

The effect of blue light on L-type calcium channel subunit mRNA expression of human retinal pigment epithelial cells cultured in vitro

作　　者：宫鑫[1] 蔡善君[1] 李海辉[1] 吕建平[1] 伍志鹏[1] 宿罡[1] 谢兵[1] 

机构地区：[1]遵义医学院附属医院眼科,563003

出　　处：《中华眼底病杂志》2013年第4期411-415,共5页Chinese Journal of Ocular Fundus Diseases

基　　金：基金项目：国家自然科学基金（81060079）;贵州省教育厅重点项目[（黔教科）2008034号]

摘　　要：目的观察蓝光照射对人视网膜色素上皮（RPE）细胞L-型钙通道mRNA表达的影响。方法体外培养的第4代人RPE细胞按随机数字表法分为无光照组、光照组、光照＋硝苯地平组和光照＋（-）BayK8644组。（2000±500）LUX蓝光照射6h，终止细胞培养24h。光照前加入硝苯地平和（-）BayK8644，浓度均为1×10^-5mmol／L。荧光定量聚合酶链（PCR）技术检测各组人RPE细胞L-型钙通道α1c亚型、α1D亚型和α1F亚型mRNA的表达。结果荧光定量PCR产物α1C、α1D、α1F和内参8-肌动蛋白的cDNA逆转录PCR扩增产物长度分别为68、157、125、186碱基对，产物的凝胶电泳结果显示扩增条带位置与以上片段相吻合。荧光定量PCR检测结果显示，光照组、光照＋硝苯地平组、光照＋（-）BayK8644组α1CtuRNA的表达均高于无光照组，差异有统计学意义（P〈0．05）；光照＋硝苯地平组、光照＋（-）BayK8644组α1CmRNA的表达均高于光照组，差异有统计学意义（P〈0．05）；光照＋（-）BayK8644组α1CmRNA的表达高于光照＋硝苯地平组，差异有统计学意义（P〈0．05）。光照组、光照＋硝苯地平组、光照＋（-）BayK8644组d1DmRNA的表达均高于无光照组，差异均有统计学意义（P〈0．05）；光照＋（-）BayK8644组α1DmRNA的表达与光照组和光照＋硝苯地平组比较，差异均有统计学意义（P〈0．05）；光照＋硝苯地平组与光照组α1DmRNA的表达比较。差异无统计学意义（P〉0．05）。光照组、光照＋硝苯地平组、光照＋（-）BayK8644组α1FmRNA的表达均高于无光照组，差异有统计学意义（P〈0．05）；光照＋硝苯地平组、光照＋（-）BayK8644组α1FmRNA的表达均高于光照组，差异有统计学意义（P〈0．05）。结论蓝光照射可引起人RPE细胞L-型钙通道中α1C、α1D及α1F亚型mRNA表达不同程度的增高；1×10^-5mmol／L硝苯地平不能对抗蓝光对RPE细胞I，型钙Objective To investigate the effect of blue light on mRNA expression of L-type calcium channel subtypes of human retinal pigment epithelial （RPE） cells in vitro. Methods The fourth-generation of human RPE cells were randomly divided into four groups including control group （no light group）, light group, light ＋ nifedipine group, and light ＋ （-） BayK8644 group. The cells were exposed to blue light （2000μ 500） lux for 6 hours, and then cultured for another 24 hours. Reverse transcription polymerase chain reaction real time （RT PCR） and fluorescence quantitative PCR technologies were used to analyze mRNA expression of L-type calcium channel subunit of cardiac subtype （ 1C or CaV1.2）, neuroendocrine subtype （ 1D or CaV1.3） and retinal subtypes （ 1F or CaV1.4） in each group. Results The length of PCR product of 1C, 1D, IF subunit and -aetin was 68, 157, 125 and 186 base pairs respectively. （1） 1C mRNA expression in light, light＋ nifedipine and light ＋ （-） BayK8644 group was higher than that in control group, the difference was statistically significant （P〈0.05）. 1C mRNA expression in light ＋nifedipine group and light ＋ （-） BayK8644 group was higher than in light group （P〈0.05）. 1C mRNA expression in light ＋ （-） BayK8644 group was higher than that in light ＋ nifedipine group （P〈 0.05）. （2） Comparing with control group, 1D mRNA expression was higher in light, light -nifedipine and light ＋（-） BayK8644 group, the difference was statistically significant （P〈0.05）. Light ＋ （-） BayK8644 group was higher than light group and light ＋ nifedipine group （P〈0.05）, light group and the light ＋ nifedipine group was not statistically significant （P〈0.05）. （3） 1F mRNA expression in light, light q- nifedipine and light ＋ （--） BayK8644 group was higher than those in control group, there was statistically significant （P〈0.05）, light ＋nifedipine group and light ＋（-） BayK8644 group was higher than

关 键 词：光刺激 视网膜色素上皮 病理生理学 钙通道 L型 

分 类 号：R77[医药卫生—眼科]