miRNA-22在脂肪基质细胞体外脂向分化过程中的表达  

Expression of miRNA-22 during adipogenic differentiation process of adipose tissue-derived stromal cells in vitro

作　　者：唐彦峰[1] 陈建霖 肖金刚[1] 张勇[3] 李晓宇[1] 田卫东[1] 刘磊[1] 

机构地区：[1]四川大学口腔疾病研究国家重点实验室,四川省成都市610041 [2]灵武市人民医院口腔科,宁夏回族自治区灵武市751400 [3]焦作煤业集团中央医院口腔科,河南省焦作市454002

出　　处：《中国组织工程研究与临床康复》2009年第33期6443-6447,共5页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：国家自然科学基金资助项目(30772422);教育部新世纪优秀人才支持计划(NCET-07-0578);四川省杰出青年学科带头人培养计划(06ZQ026-008)~~

摘　　要：背景:脂肪基质细胞在特定条件下可以向脂肪细胞分化,然而,脂肪基质细胞脂向分化的分子机制仍不明确!目的:明确miRNA在脂肪基质细胞脂向分化过程中是否发挥作用及其可能机制。设计、时间及地点:观察对照试验,于2007-10/2008-03在四川大学口腔疾病研究国家重点实验室完成。材料:出生30d健康雄性SD幼鼠8只,取出脂肪,采用密度梯度离心结合贴壁培养法获得了纯度高的脂肪基质细胞。方法:采用脂向诱导液对第3代的脂肪基质细胞进行诱导培养。建立脂肪基质细胞体外脂向分化细胞模型,采用miRNA芯片技术筛选脂肪基质细胞向脂肪细胞方向分化的相关miRNA,使用实时定量聚合酶链反应技术对芯片结果进行验证。主要观察指标:①细胞形态学。②PPARγ免疫细胞化学染色和油红O染色。③miRNA芯片结果。④实时定量聚合酶链反应结果。结果:①经显微镜下观察、油红O染色及PPARγ免疫细胞化学染色检测证实脂肪基质细胞已经脂向分化。②miRNA芯片发现诱导组中miRNA-22的表达量呈下降趋势。③实时定量聚合酶链反应结果证实miRNA-22的表达量与对照组相比显著减少(P<0.05)。结论:miRNA-22在脂肪基质细胞脂向分化后的表达有显著降低,提示miRNA-22可能参与脂肪基质细胞的脂肪分化调控过程,生物信息学分析提示,该过程可能通过对其靶基因MAPK7(ERK5)的调控而实现。BACKGROUND： Adipose derived stromal cells （ADSCs） can differentiate to lipocytes under specific conditions, however, the detailed regulatory mechanisms of ADSCs adipogenic differentiation have not yet completely understood. OBJECTIVE： To definite the acts and the possible mechanism of miRNA in the adipogenic differentiation process of ADSCs. DESIGN, TIME AND SETTING： An observation controlled experiment was performed in the State Key Laboratory of Oral Diseases, Sichuan University between October 2007 and March 2008. MATERIALS： Eight male SD neonatal rats within 30 birth days were used to take out fat tissues and to obtain highly purified ADSCs with density gradient centrifugation and adherent culture methods. METHODS： The third generation of ADSCs were induced into adipogenic differentiation with the inductive medium. The in vitro model of the adipogenic differentiation of ADSCs was established, and the miRNA gene chip technology was applied to screen the correlated miRNA during the ADSCs adipogenesis. Real-time quantitative PCR was used to verify the miRNA result. MAIN OUTCOME MEASURES： ①Cellular morphology; ②Oil red O and immunocytochemical detection of PPARγ; ③ miRNA-22 chip; ④real-time quantitative PCR. RESULTS： ①The adipogenic differentiation of ADSCs was confirmed by microscopy, staining with Oil red O and immunocytochemical detection of PPARγ. ②The miRNA-22 expression had a declining trend in the induced group discovered in the miRNA result. ③The result of real-time quantitative PCR showed that the miRNA-22 expression in the induction group was significantly lower than that in the control group （P 〈 0.05）. CONCLUSION： The miRNA-22 expression have a remarkable decline in the adipogenic differentiation process of ADSCs, which indicates that miRNA-22 may be involved in the regulation process of adipogenic differentiation of ADSCs. Bioinformatics analysis shows that the adipogenic differentiation process is possibly carried out by regulating MAPK7 （ERK5）, which is

关 键 词：脂肪基质细胞 miRNA-22 实时定量聚合酶链反应技术 成脂分化 

分 类 号：R318[医药卫生—生物医学工程]