CAJ | 학술논문

目的探讨miR-181a和miR-181b靶向调控bcl-2及其在年龄相关性白内障发生中的作用机制.方法临床病例系列研究.对2014年10～12月在中国医科大学附属第四医院眼科利用生物信息学软件,预测miR-181a和miR-181b可能调控的靶基因;利用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miR-181a、miR-181b及其预测靶基因的表达;利用Lipofectamine 2000瞬时转染miR-181a和miR-181b mimic和inhibitor,调节人晶状体上皮细胞中miR-181a和miR-181b的表达水平,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.结果生物信息学软件预测到bcl-2可能为miR-181a和miR-181b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-181a和miR-181b的表达均显著升高,而bcl-2的表达显著降低.miR-181a和miR-181b mimic转染组miR-181a和miR-181b的表达显著升高,bcl-2的表达显著降低;miR-181a和miR-181b inhibitor转染组,miR-181a和miR-181b的表达显著降低,bcl-2的表达显著升高.差异均有统计学意义(P＜0.01).结论 miR-181a和miR-181b在年龄相关性白内障晶状体组织中呈高表达,其可能靶向bcl-2,对其负性调控,从而影响白内障的发病过程.
목적 탐토miR-181a화miR-181b파향조공bcl-2급기재년령상관성백내장발생중적작용궤제.방법 림상병례계렬연구.대2014년10～12월재중국의과대학부속제사의원안과이용생물신식학연건,예측miR-181a화miR-181b가능조공적파기인;이용Real time q-PCR방법,검측년령상관성백내장환자정상체전낭막화정상인안정상체전낭막miR-181a、miR-181b급기예측파기인적표체;이용Lipofectamine 2000순시전염miR-181a화miR-181b mimic화inhibitor,조절인정상체상피세포중miR-181a화miR-181b적표체수평,이용Real time q-PCR방법험증전염효솔,병검측예측파기인적표체변화.결과 생물신식학연건예측도bcl-2가능위miR-181a화miR-181b적파기인.여대조조상비,년령상관성백내장환자정상체전낭막조,miR-181a화miR-181b적표체균현저승고,이bcl-2적표체현저강저.miR-181a화miR-181b mimic전염조miR-181a화miR-181b적표체현저승고,bcl-2적표체현저강저;miR-181a화miR-181b inhibitor전염조,miR-181a화miR-181b적표체현저강저,bcl-2적표체현저승고.차이균유통계학의의(P＜0.01).결론 miR-181a화miR-181b재년령상관성백내장정상체조직중정고표체,기가능파향bcl-2,대기부성조공,종이영향백내장적발병과정.
Objective To study the mechanism of miR-181a and miR-181b targeted bcl-2 in the pathogenesis of age-related cataract.Methods Biological information softwares were used to predict potential target gene of miR-181a and miR-181b.Real time q-PCR was used to measure the expression of miR-181a,miR-181b and potential target genes between the anterior lens capsules of age-related cataract and normal anterior lens capsule specimens.MiR-181a and miR-181b mimic and inhibitor were transfected into SRA01/04 cells using Lipofectamine 2000 to regulate the expression of miR-181a and miR-181b,and then real time q-PCR was used to assess transfection efficiency and the expression of potential target genes.Results Bcl-2 was predicted as the potential target gene of miR-181a and miR-181b using three different biological information softwares.Compared with controls,the expression of miR-181a and miR-181b was significantly higher in the anterior lens capsules of age-related cataract.On the contrary,the expression of bcl-2 was decreased in the same tissue specimens.The relative expression of miR-181a and miR-181b in lens epithelial cells transfected with miR-181a and miR-181b mimic was increased,whereas was decreased in cells transfected with miR-181a and miR-181b inhibitor.The expression of bcl-2 transfected with miR-181a and miR-181b mimic was reduced,while increased in miR-181a and miR-181b inhibitor group.Each result was statistically significant (P ＜0.01).Conclusions High expression of miR-181a and miR-181b is detected in anterior lens capsule of age-related cataract.MiR-181a and miR-181b may play an important role in the pathogenesis of age-related cataract by targeting bcl-2.