胰岛素抵抗小鼠血清微小RNA的芯片及信息学分析

Chip and informatic analysis of serum microRNA levels in insulin-resistant mice

目的:探讨胰岛素抵抗小鼠血清水平微小RNA(microRNAs)变化图谱及相关microRNAs的可能作用机制。方法:以高脂饲料喂养昆明小鼠,制备胰岛素抵抗模型;以微阵列芯片技术分析胰岛素抵抗小鼠及正常小鼠血清水平microRNAs变化图谱并以实时荧光定量PCR进行验证;以miRanda数据库对差异microRNAs进行靶点基因预测;以miRBase数据库获得与胰岛素抵抗相关的microRNAs序列;基于microRNAs序列,应用STRING在线分析工具(http://string.embl.de/)预测蛋白质相互作用关系。结果:胰岛素抵抗小鼠血清中miR-125、miR-126、miR-143、miR-30a、miR-199a、miR-127、miR-184、miR-30e、miR-134、miR-195、miR-206、miR-429、miR-212、miR-362、miR-382、miR-154和miR-466h表达显著上调,miR-211、miR-504、miR-877和miR-1930显著下调。与胰岛素抵抗密切相关的miR-143可以结合脂肪量及肥胖相关蛋白(FTO)的3’-UTR,并且FTO与Rpgrip1l、Tmem18、Mc4r、Npy、Hhex、Tcf712、Cdkal1、Slc30a8、Igf2bp2和Thada相互作用。结论:在高脂饮食诱导的胰岛素抵抗小鼠血清中有21种microRNAs相比正常小鼠出现了显著变化,表达显著上调的有17个。应用在线分析工具发现与胰岛素抵抗密切相关的miR-143可以调节FTO蛋白表达,并且FTO与另外10种与糖尿病发生发展相关的蛋白相互作用,这将有助于对胰岛素抵抗机制的全面了解。

AIM： To study the microRNA profiling in the serum of insulin-resistant mice and the mechanism of insulin resistance induced by related microRNAs. METHODS： A high-fat diet was used to induce insulin resistance model in KM mice. The microRNA profiling in serum of insulin-resistant and normal mice was analyzed by microarray chip and were validated by real-time PCR. miRanda data base was used to forecast target genes. miRBase was used to obtain the sequences of related microRNAs,based on which protein interactions were predicted using the online analytical tool STRING.RESULTS： In serum of insulin-resistant mice,the expression of miR-125,miR-126,miR-143,miR-30 a,miR-199 a,miR-127,miR-184,miR-30 e,miR-134,miR-195,miR-206,miR-429,miR-212,miR-362,miR-382,miR-154 and miR-466 h was significantly up-regulated. miR-211,miR-504,miR-877 and miR-1930 were significantly down-regulated.miR-143 associated with insulin resistance was able to bind to 3＇-UTR of fat mass and obesity-associated protein（ FTO）,and FTO was found to interact with Rpgrip1 l,Tmem18,Mc4 r,Npy,Hhex,Tcf712,Cdkal1,Slc30a8,Igf2bp2 and Thada. CONCLUSION： Twenty-one microRNAs in the serum of insulin-resistant mice induced by a high-fat diet are significantly different from those of normal mice,in which 17 kinds were significantly up-regulated. miR-143 closely related to insulin resistance is able to regulate FTO protein expression,which interacts with other 10 proteins associated with the occurrence and development of diabetes. The results are also useful for further study of the molecular mechanisms in insulin resistance.