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目的探讨miR-181a在胸腺增龄性萎缩过程中的表达,及其与靶基因ZFP36l2的相互作用关系,阐明胸腺增龄性萎缩过程分子调节机制。方法 C57BL/6小鼠分为1月龄组、9月龄组、15月龄组,每组各6只。取出胸腺并分离细胞,经CD45抗体染色,LS柱吸附洗脱筛选出胸腺细胞。实时荧光定量PCR法检测胸腺细胞中miR-181a与靶基因ZFP36l2随年龄增长的表达变化趋势,并进行相关性分析。体外应用HEK293细胞转染荧光素酶报告载体,48 h后检测自发荧光值。结果随年龄增长,miR-181a呈现表达下调趋势(P<0.05),而靶基因ZFP36l2则呈现出表达上升趋势(P<0.05)。对miR-181a与ZFP36l2表达量进行相关性分析,发现两者呈负相关性(P=0.04)。体外细胞转染荧光素酶报告基因证实,miR-181a与ZFP36l2 3’UTR区域结合(P<0.05)。结论 miR-181a与靶基因ZFP36l2与胸腺增龄性萎缩过程密切相关,是调节胸腺细胞功能的重要因子。
Objective To investigate the expression of miR-181a in the process of atrophic thymus and its interaction with target gene ZFP36l2, and to elucidate the molecular regulation mechanism of the thymus atrophy process. Methods C57BL / 6 mice were divided into 1-month-old, 9-month-old and 15-month-old groups, 6 in each. The thymus was removed and the cells were detached, stained with CD45 antibody, and the thymus cells were screened by LS column. Real-time fluorescence quantitative PCR was used to detect the change trend of miR-181a and target gene ZFP36l2 in thymocytes with age, and the correlation was analyzed. HEK293 cells were transfected with luciferase reporter vector in vitro, and autofluorescence was detected 48 h later. Results With the increase of age, miR-181a expression was down-regulated (P <0.05), while target gene ZFP36l2 showed an up-regulated expression (P <0.05). Correlation analysis between miR-181a and ZFP36l2 expression showed a negative correlation (P = 0.04). In vitro cell transfection with luciferase reporter gene confirmed that miR-181a bound to the ZFP36l2 3’UTR region (P <0.05). Conclusion miR-181a and target gene ZFP36l2 are closely related to the process of atrophic thymus, which is an important factor in the regulation of thymocyte function.