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目的探讨microRNAs对脂肪细胞分化过程的调节作用分子机制及其对脂肪特异性基因——脂肪型脂肪酸结合蛋白(fatty acid binding protein,FABP4)表达的影响。方法用microRNAs微阵列分析法筛选和鉴定3T3-L1脂肪细胞分化相关性microRNAs,构建脂肪细胞分化相关性micro-RNAs的高表达质粒,通过脂质体介导转染3T3-L1前体脂肪细胞,观察脂肪相关性microRNAs对脂肪细胞分化过程的影响,Westernblot和RT-PCR分别检测FABP4蛋白及其mR-NA在脂肪细胞分化过程中的表达变化。结果3T3-L1脂肪细胞分化过程中microRNA表达谱发生明显改变,其中35个microRNAs下调,miR-24最明显;17个microRNAs上调,miR-21最明显;用不同脂肪相关性microRNAs转染3T3-L1前体脂肪细胞并高表达后,发现miR-24明显抑制脂肪细胞的分化与成熟,而miR-21则无影响;miR-24明显抑制FABP4蛋白表达,但对其mRNA无影响,miR-21对FABP4的蛋白和mRNA表达都没有影响。结论3T3-L1脂肪细胞分化过程中存在脂肪细胞分化相关性microRNAs;miR-24脂肪细胞分化过程及其脂肪特异蛋白FABP4表达有重要的调节作用。
Objective To investigate the molecular mechanism of microRNAs regulation on adipocyte differentiation and its effect on the expression of fat-specific fatty acid binding protein (FABP4). Methods 3T3-L1 adipocyte differentiation-related microRNAs were screened and identified by microRNAs microarray analysis to construct high expression plasmids of adipocyte differentiation-associated microRNAs. Liposome-mediated transfection of 3T3-L1 precursor adipocytes, Observe the effect of fat-related microRNAs on adipocyte differentiation. Western blot and RT-PCR were used to detect the expression of FABP4 protein and mR-NA in adipocyte differentiation. Results The microRNA expression profiles of 3T3-L1 adipocytes were significantly changed during differentiation. Among them, 35 microRNAs were down-regulated and miR-24 was the most obvious. 17 microRNAs were up-regulated and miR-21 was the most obvious. Transfection of 3T3-L1 with different adipose- Pre-adipocytes and high expression, we found miR-24 significantly inhibited adipocyte differentiation and maturation, and miR-21 had no effect; miR-24 significantly inhibited FABP4 protein expression, but no effect on its mRNA, miR-21 pairs FABP4 protein and mRNA expression had no effect. Conclusion There are adipocyte differentiation-related microRNAs during the differentiation of 3T3-L1 adipocytes. The differentiation of miR-24 adipocytes and the expression of FABP4 are important regulatory roles.