脂肪基质细胞脂向分化过程中MicroRNA-21的表达变化  

Changes in MicroRNA-21 Expression during Adipogenic Differentiation Of Adipose Tissue-derived Stromal Cells

作　　者：王伟新[1,2] 张勇[1] 李晓宇[1] 龙洁[1] 田卫东[2] 汤炜[2] 王杭[2] 郑晓辉[1] 刘磊[2] 

机构地区：[1]四川大学口腔疾病研究国家重点实验室,四川成都610041 [2]四川大学华西口腔医院口腔颌面外科,四川成都610041

出　　处：《现代生物医学进展》2009年第23期4404-4406,4459,共4页Progress in Modern Biomedicine

基　　金：国家自然科学基金(30772422);教育部新世纪优秀人才支持计划(JS20071108506525);四川省杰出青年学科带头人培养计划(06ZQ026-008);四川省支撑计划项目(2009FZ0074)

摘　　要：目的:了解脂肪基质细胞(ADSCs)在脂向分化过程中MicroRNA-21(miRNA-21)表达变化情况。方法:从8只出生30d雄性体健SD幼鼠体内取出脂肪,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,采用脂向诱导液对第3代的脂肪基质细胞进行诱导培养,并以未诱导的细胞为对照。成脂诱导培养液[DMEM培养液中加入地塞米松(1μmol.L-1)、3-异丁基-1-甲基黄嘌呤(0.5mmol.L-1)、胰岛素(10mg.L-1)]。应用microRNA芯片技术检测脂肪基质细胞脂向诱导后7d和未诱导microRNA-21的表达差异。采用实时定量PCR检测microRNA-21在脂肪基质细胞脂向分化前后表达量变化。结果: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,miRNA芯片及实时定量PCR结果均表明miRNA-21在脂肪基质细胞体外脂向分化过程中表达显著下调。结论:miRNA-21在脂肪基质细胞脂向分化后的表达有显著降低,可能参与调控ADSCs的脂向分化过程。Objective： To explore the differential expression of microRNA-16 during adipogenic differentiation of adipose derived stromalcells （ADSCs）. Methods： The adipose was removed from eight 30-day-old Sprague-Dawle SD rats, and was treated with density gradient centrifugation and adherent culture to obtain ADSCs. The third generation cells were induced and cultured into adipogenic differentiation with Adipogenic-differentiated culture medium [DMEM includingdexamethasone （11μmol·L^-1）, 3-isobutyl-1-methyl xanthine （0.5 mmol.L^-1）-land insulin （10 mg.L^-1 ）, and the non-induced cells were served as the control group. Results： The adipogenic differentiation of ADSCs was confirmed by microscopy, staining with Oil red 0 and immunocytochemical detection of PPARγ The miRNA-21 was identified by the miRNA chip technology as being down -regulated during adipogenic differentiation of ADSCs. Real -time PCR analysis confirmed the decreased expression of miRNA-21. Conclusion： The miRNA-21 expression will significantly decrease in the adipogenic differentiation process of ADSCs, which maybe involved in the regulation process of adipogenic differentiation of ADSCs.

关 键 词：MICRORNA-21 脂肪基质细胞 脂向分化 microRNA芯片 实时定量PCR 

分 类 号：Q95-3[生物学—动物学] Q819