CAJ | 학술논문

目的探讨miR-204靶向调控Bcl-2家族的机制及其在年龄相关性白内障发病过程中的作用.方法临床实验研究.于2014年10～12月在中国医科大学附属第四医院眼科,收集年龄相关性白内障患者晶状体前囊膜标本36例为实验组,选取新鲜人眼透明晶状体前囊膜标本12例为对照组.利用生物信息学软件,预测miR-204可能调控的Bcl-2家族关键基因;应用Real time q-PCR方法,检测年龄相关性白内障患者晶状体前囊膜和正常人眼晶状体前囊膜间miR-204及其预测靶基因的表达;利用Lipofectamine 2000瞬时转染miR-204 mimic和inhihitor,分别上调和下调人晶状体上皮细胞中miR-204的表达,利用Real time q-PCR方法验证转染效率,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Bcl-2家族中bcl-2、mcl-1可能为miR-204的靶基因.与对照组相比,年龄相关性白内障患者晶状体前囊膜组,miR-204的表达显著升高,bcl-2、mcl-1的表达显著降低.miR-204 mimic转染组,miR-204的表达显著升高,bcl-2、mcl-1的表达显著降低;miR-204 in-hibitor转染组,miR-204的表达显著降低,bcl-2、mcl-1的表达显著升高.差异均有统计学意义(P＜0.01).结论 miR-204在年龄相关性白内障晶状体组织中呈高表达,miR-204可能通过靶向调控Bcl-2家族成员bcl-2、mcl-1的表达,在年龄相关性白内障发病过程中发挥重要角色,miR-204可能成为白内障非手术治疗的新靶点.
목적 탐토miR-204파향조공Bcl-2가족적궤제급기재년령상관성백내장발병과정중적작용.방법 림상실험연구.우2014년10～12월재중국의과대학부속제사의원안과,수집년령상관성백내장환자정상체전낭막표본36례위실험조,선취신선인안투명정상체전낭막표본12례위대조조.이용생물신식학연건,예측miR-204가능조공적Bcl-2가족관건기인;응용Real time q-PCR방법,검측년령상관성백내장환자정상체전낭막화정상인안정상체전낭막간miR-204급기예측파기인적표체;이용Lipofectamine 2000순시전염miR-204 mimic화inhihitor,분별상조화하조인정상체상피세포중miR-204적표체,이용Real time q-PCR방법험증전염효솔,병검측예측파기인적표체변화.결과 3충불동적생물신식학연건예측도Bcl-2가족중bcl-2、mcl-1가능위miR-204적파기인.여대조조상비,년령상관성백내장환자정상체전낭막조,miR-204적표체현저승고,bcl-2、mcl-1적표체현저강저.miR-204 mimic전염조,miR-204적표체현저승고,bcl-2、mcl-1적표체현저강저;miR-204 in-hibitor전염조,miR-204적표체현저강저,bcl-2、mcl-1적표체현저승고.차이균유통계학의의(P＜0.01).결론 miR-204재년령상관성백내장정상체조직중정고표체,miR-204가능통과파향조공Bcl-2가족성원bcl-2、mcl-1적표체,재년령상관성백내장발병과정중발휘중요각색,miR-204가능성위백내장비수술치료적신파점.
Objective To study the mechanism of miR-204 targeted Bcl-2 family in the pathogenesis of age-related cataract.Methods Clinical and experimental research.Thirty-six fresh specimens of anterior lens capsule of age-related cataract were obtained as experimental group from cataract patients at the Fourth Affiliated Hospital of china Medical University between October 2014 and December 2014.Twelve normal anterior lens capsule specimens were obtained as control group.Biological information softwares were used to predict potential target genes of miR-204 in Bcl-2 family.Real time q-PCR was used to measure the expression of miR-204 and potential target genes between the anterior lens capsules of age-related cataract and normal anterior lens capsule specimens.MiR-204 mimic and inhibitor were transfected into SRA01/04 cells using Lipofectamine 2000 to regulate the expression of miR-204,and then real time q-PCR was used to assess transfection efficiency and the expression of potential target genes.Results Bcl-2 and mcl-1 were predicted as the potential target genes of miR-204 using three different biological information softwares.Compared with controls,the expression of miR-204 was significantly higher in the anterior lens capsules of age-related cataract.On the contrary,the expression of bcl-2 and mcl-1 was decreased in the same tissue specimens.The relative expression of miR-204 in lens epithelial cells transfected with miR-204 mimic was increased,whereas was decreased in cells transfected with miR-204 inhibitor.The expression of bcl-2 and mcl-1 transfected with miR-204 mimic was reduced,while increased in miR-204 inhibitor group.Each result was statistically significant (P ＜0.01).Conclusions High expression of miR-204 is detected in anterior lens capsule of age-related cataract.MiR-204 may play a critical role in the pathogenesis of age-related cataract by targeting bcl-2 and mcl-1 in Bcl-2 family.MiR-204 might be a new target for nonoperative treatment of cataract.