microRNA-215在糖尿病肾病小鼠肾组织中的表达及意义  

Role of microRNA-215 in nephropathy of type 2 diabetic db/db mice

作　　者：庞琦[1] 牟娇[1] 郭艳红[1] 陈吉刚[1] 曾薇[1] 黄拥军[1] 张均[1] 钱丹[1] 冯兵[1] 

机构地区：[1]第三军医大学新桥医院肾内科,重庆400037

出　　处：《中华肾脏病杂志》2012年第4期305-311,共7页Chinese Journal of Nephrology

基　　金：国家自然科学基金（2009A214）

摘　　要：目的探讨microRNA-215（miR-215）在糖尿病肾病（DN）小鼠。肾组织中的表达变化规律及在DN发病中的作用。方法选择4周龄的2型糖尿病肾病db／db小鼠（实验组）和db／m小鼠（对照组），采用实时荧光定量PCR法动态检测8、12及16周龄时肾组织miR-215的表达变化；实时荧光定量PCR和Western印迹法、免疫组化法测定连环蛋白B互动蛋白1（CTNNBIP1）的mRNA及蛋白的表达；双荧光素酶报告法确证miR-215对CTNNBIP1表达的直接调控作用。结果（1）随着周龄的增加，db／db小鼠肾小球逐渐肥大、节段性系膜细胞增生和系膜基质积聚。（2）与同周龄的db／m小鼠比较，8、12及16周龄的db／db小鼠体质量（BW）、血糖（Glu）及24h尿白蛋白排泄量（UAE）均显著增加（均P〈0．05）。（3）随着周龄的增加，db／db小鼠肾脏组织miR-215表达显著高于同周龄的db／m小鼠（P〈0．05）。（4）与同周龄的db／m小鼠比较，db／db小鼠肾脏组织CTNNBIPlmRNA和蛋白均显著降低（均P〈0．05）。（5）应用双荧光素酶报告法证实，mi-215可显著抑制CTNNBIPl的表达（P〈0．01）。结论miR-215表达上调可能通过抑制CTNNBIPl的表达参与DN的发生、发展。Objective To investigate the renal expression changes of microRNA-215 （miR-215） and its role in diabetic nephropathy of type 2 diabetic db/db mice. Methods Four- week-old diabetic db/db mice and normal control group non-diabetic db/m mice were selected. Real-time PCR was used to detect the relative level of miR-215 at the age of 8, 12 and 16 weeks. Catenin beta interacting protein 1 （CTNNBIPI） mRNA and protein level were measured by real- time PCR, Western blotting and immunohistochemisty. A lueiferase reporter assay was used to determine whether CTNNBIP1 was a direct target of miR-215. Results （1）With the growth of db/db mice, the major pathological characteristics of kidney ineluded glomerular hypertrophy, segmental mesangial ceils proliferation and mesangial matrix expansion. （2）Compared with the db/m mice, the db/db mice of 8, 12 and 16 weeks showed obvious increase in body weight（BW）, blood glueose （Glu） and 24 hour urinary albumin excretion （UAE） （P〈0.05, respectively）. （3）Compared with the dh/m mice, special miR-215 was highly expressed in the kidney of db/db mice and was up-regulated signifieantly according to the development of DN （P〈0.05）. （4）The mRNA and protein expression of CTNNBIP1 of kidney were consistently down-regulated in db/db mice than those incontrols （P〈0.05, respectively）. （5）By luciferase reporter, miR-215 could negatively regulate CTNNBIP1 gene by targeting its 3＇-UTR sequence （P〈0.01）. Conclusion High expression level of miR-215 plays a potential role in the initiation and progression of DN by down-regulating the expression of CTNNBIP1.

关 键 词：糖尿病肾病 微RNAS Β连环素 互动蛋白1 双荧光素酶报告 

分 类 号：R587.2[医药卫生—内分泌]