CAJ | 학술논문

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β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24 h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CTNNBIP1 mRNA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,miR-215可显著抑制CTNNBIP1的表达(P＜0.01).结论 miR-215表达上调可能通过抑制CTNNBIP1的表达参与DN的发生、发展.
목적 탐토microRNA-215( miR-215)재당뇨병신병(DN)소서신조직중적표체변화규률급재DN발병중적작용.방법 선택4주령적2형당뇨병신병db/db소서(실험조)화db/m소서(대조조),채용실시형광정량PCR법동태검측8、12급16주령시신조직miR-215적표체변화；실시형광정량PCR화Western인적법、면역조화법측정련배단백β호동단백1( CTNNBIP1)적mRNA급단백적표체；쌍형광소매보고법학증miR-215대CTNNBIP1표체적직접조공작용.결과 (1)수착주령적증가,db/db소서신소구축점비대、절단성계막세포증생화계막기질적취.(2)여동주령적db/m소서비교,8、12급16주령적db/db소서체질량(BW)、혈당( Glu)급24 h뇨백단백배설량(UAE)균현저증가(균P＜0.05).(3)수착주령적증가,db/db소서신장조직miR-215표체현저고우동주령적db/m소서(P＜0.05).(4)여동주령적db/m소서비교,db/db소서신장조직CTNNBIP1 mRNA화단백균현저강저(균P＜0.05).(5)응용쌍형광소매보고법증실,miR-215가현저억제CTNNBIP1적표체(P＜0.01).결론 miR-215표체상조가능통과억제CTNNBIP1적표체삼여DN적발생、발전.
Objective To investigate the renal expression changes of microRNA-215(miR-215) and its role in diabetic nephmpathy of type 2 diabetic db/db mice. Methods Fourweek-old diabetic db/db mice and norml 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 (CTNNBIP1) mRNA and protein level were measured by realtime 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 included glomerular hypertrophy,segmental mesangial cells 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 glucose (Glu) and 24 hour urinary albumin excretion (UAE) (P＜0.05,respectively).(3)Compared with the db/m mice,special miR-215 was highly expressed in the kidney of db/db mice and was up-regulated significantly according to the development of DN (P＜0.05).(4)The mRNA and protein expression of CTNNBIPl of kidney were consistently down-regulated in db/db mice than those in controls (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 CTNNBIPl.