CAJ | 학술논문

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24 h.光照前加入硝苯地平和(-)BayK8644,浓度均为1×10-5 mmol/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β-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α1C mRNA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α1C mRNA的表达均高于光照组,差异有统计学意义(P＜0.05)；光照+(-)BayK8644组α1C mRNA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α1D mRNA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α1D mRNA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α1D mRNA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α1F mRNA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α1F mRNA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；1×10-5 mmol/L硝苯地平不能对抗蓝光对RPE细胞L-型钙通道作用,而1×10-5 mmol/L(-)Bay K8644可以增加以上3个亚基的表达.
목적 관찰람광조사대인시망막색소상피(RPE)세포L-형개통도mRNA표체적영향.방법 체외배양적제4대인RPE세포안수궤수자표법분위무광조조、광조조、광조+초분지평조화광조+(-)BayK8644조.(2000±500) Lux람광조사6h,종지세포배양24 h.광조전가입초분지평화(-)BayK8644,농도균위1×10-5 mmol/L.형광정량취합매련(PCR)기술검측각조인RPE세포L-형개통도α1C아형、α1D아형화α1F아형mRNA적표체.결과 형광정량PCR산물α1C、α1D、α1F화내삼β-기동단백적cDNA역전록PCR확증산물장도분별위68、157、125、186감기대,산물적응효전영결과현시확증조대위치여이상편단상문합.형광정량PCR검측결과현시,광조조、광조+초분지평조、광조+(-)BayK8644조α1C mRNA적표체균고우무광조조,차이유통계학의의(P＜0.05)；광조+초분지평조、광조+(-)BayK8644조α1C mRNA적표체균고우광조조,차이유통계학의의(P＜0.05)；광조+(-)BayK8644조α1C mRNA적표체고우광조+초분지평조,차이유통계학의의(P＜0.05).광조조、광조+초분지평조、광조+(-)BayK8644조α1D mRNA적표체균고우무광조조,차이균유통계학의의(P＜0.05)；광조+(-)BayK8644조α1D mRNA적표체여광조조화광조+초분지평조비교,차이균유통계학의의(P＜0.05)；광조+초분지평조여광조조α1D mRNA적표체비교,차이무통계학의의(P＞0.05).광조조、광조+초분지평조、광조+(-)BayK8644조α1F mRNA적표체균고우무광조조,차이유통계학의의(P＜0.05)；광조+초분지평조、광조+(-)BayK8644조α1F mRNA적표체균고우광조조,차이유통계학의의(P＜0.05).결론 람광조사가인기인RPE세포L-형개통도중α1C、α1D급α1F아형mRNA표체불동정도적증고；1×10-5 mmol/L초분지평불능대항람광대RPE세포L-형개통도작용,이1×10-5 mmol/L(-)Bay K8644가이증가이상3개아기적표체.
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 (2000± 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,1F subunit and-actin 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 +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 light group (P＜0.05),light + nifedipine group and the light + (-) BayK8644 group was not statistically significant (P＞0.05).Conclusions The human RPE cells mRNA expression of L-type calcium channel 1C,1D and 1F subunit was increased after exposing to blue light.Application of the 1 × 10-s mmol/L (-)BayK8644 can increase mRNA expression of 1C,1D and 1F subunit.