CAJ | 학술논문

目的：探讨microRNA let-7a靶向调控Caspase家族的机制及其在年龄相关性白内障发病过程中的作用。方法收集2014年7~9月在中国医科大学附属第四医院(以下简称“我院”)眼科诊断为年龄相关性白内障(排除其他眼部疾病)的患者晶状体前囊膜标本30例，作为实验组；同时收集我院眼科眼库提供的新鲜人眼透明晶状体前囊膜标本15例，作为对照组。利用生物信息学软件预测let-7a可能调控的Caspase家族中的关键基因；采用实时荧光定量PCR (Real time q-PCR)检测实验组和对照组let-7a及其预测靶基因的表达；利用Lipofectamine 2000瞬时转染let-7a mimic和inhibitor，调节人晶状体上皮细胞中let-7a的表达，利用Real time q-PCR法验证转染效率，并检测预测靶基因的表达变化。结果3种不同的生物信息学软件预测到Caspase家族中Caspase-3、Caspase-8、Caspase-9可能为let-7a的靶基因。与对照组比较，实验组let-7a的表达显著降低，Caspase-3、Cas-pase-8、Caspase-9的表达显著升高。let-7a mimic转染组，let-7a的表达显著升高，Caspase-3、Caspase-8、Caspase-9的表达显著降低；let-7a inhibitor转染组，let-7a的表达显著降低，Caspase-3、Caspase-8、Caspase-9的表达显著升高；差异均有高度统计学意义(P<0.01)。结论let-7a可能通过靶向调控Caspase家族成员Caspase-3、Caspase-8、Caspase-9的表达，在年龄相关性白内障发病过程中发挥重要作用，let-7a可能成为白内障非手术治疗的新靶点。
목적：탐토microRNA let-7a파향조공Caspase가족적궤제급기재년령상관성백내장발병과정중적작용。방법수집2014년7~9월재중국의과대학부속제사의원(이하간칭“아원”)안과진단위년령상관성백내장(배제기타안부질병)적환자정상체전낭막표본30례，작위실험조；동시수집아원안과안고제공적신선인안투명정상체전낭막표본15례，작위대조조。이용생물신식학연건예측let-7a가능조공적Caspase가족중적관건기인；채용실시형광정량PCR (Real time q-PCR)검측실험조화대조조let-7a급기예측파기인적표체；이용Lipofectamine 2000순시전염let-7a mimic화inhibitor，조절인정상체상피세포중let-7a적표체，이용Real time q-PCR법험증전염효솔，병검측예측파기인적표체변화。결과3충불동적생물신식학연건예측도Caspase가족중Caspase-3、Caspase-8、Caspase-9가능위let-7a적파기인。여대조조비교，실험조let-7a적표체현저강저，Caspase-3、Cas-pase-8、Caspase-9적표체현저승고。let-7a mimic전염조，let-7a적표체현저승고，Caspase-3、Caspase-8、Caspase-9적표체현저강저；let-7a inhibitor전염조，let-7a적표체현저강저，Caspase-3、Caspase-8、Caspase-9적표체현저승고；차이균유고도통계학의의(P<0.01)。결론let-7a가능통과파향조공Caspase가족성원Caspase-3、Caspase-8、Caspase-9적표체，재년령상관성백내장발병과정중발휘중요작용，let-7a가능성위백내장비수술치료적신파점。
Objective To study the mechanism of microRNA let-7a targeted Caspase family in the pathogenesis of age-related cataract. Methods 30 fresh specimens of anterior lens capsule of age-related cataract were obtained as experi-mental group from cataract patients with no other eye diseases during phacoemulsification at the Department of Oph-thalmology, the Fourth Affiliated Hospital of China Medical University (“our hospital”for short) form July to September 2014. 15 normal anterior lens capsule specimens were obtained as control group from eye bank of Department of Oph-thalmology in our hospital. Biological information softwares were used to predict potential target genes of let-7a in Cas-pase family. Real time q-PCR was used to measure the expression of let-7a and potential target genes between the ex-perimental group and the control group. Let-7a mimic and inhibitor were transfected into SRA01/04 cells using Lipo-fectamine 2000 to regulate the expression of let-7a, and then real time q-PCR was used to assess transfection efficien-cy and the expression of potential target genes. Results Caspase-3, Caspase-8 and Caspase-9 were predicted as the potential target genes of let-7a using three different biological information softwares. Compared with control group, the expression of let-7a was significantly lower in the ex-perimental group. On the contrary, the expression of Caspase-3, Caspase-8 and Caspase-9 increased in the same tissue specimens. The relative expression of let-7a in lens epithelial cells transfected with let-7a mimic increased, whereas decreased in cells transfected with let-7a inhibitor. The expression of Caspase-3, Caspase-8 and Caspase-9 transfected with let-7a mimic were reduced, while increased in let-7a inhibitor group, the differences were statistically significant (P < 0.01). Conclusion Let-7a may play a critical role in the pathogenesis of age-related cataract by targeting caspase-3, Caspase-8 and Caspase-9 in Caspase family. Let-7a might be a new target for nonop-erative treatment of cataract.