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.