Yu Yang, Li Xue, Liu Wei-Jiang, Chen Xiu-Hui, Liu Yuan-Lin, Zhang Wei, Zhang Yi
School of Postgraduate, Anhui Medical University, Hefei 230032, Anhui Province, China; Department of Cell Biology, Institute of Basic Medical Sciences, Academy of Military Medical Sciences, Beijing 100850, China.
Department of Cell Biology, Institute of Basic Medical Sciences, Academy of Military Medical Sciences, Beijing 100850, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2017 Apr;25(2):541-548. doi: 10.7534/j.issn.1009-2137.2017.02.042.
To explore the effect of MicroRNA-146b-5p (miR-146b-5p) on the mouse bone essence derived MSC adipogenic differentiation.
MSC were isolated from bone essence of C57BL/6 mice. The expression level of miR-146b-5p in the process of adipogenic differentiation of MSC was detected by q-PCR; the role of miR-146b-5p mimics or inhibitors in the process of mouse bone essence derived MSC adipogenic differentiation was analyzed through oil red staining the expression of C/EBPα and PPARγ after cultured for 14 days was detected by q-PCR; the protein level of PPARγ after miR-146b-5p transfection was detected by Western blot.
The MSC were successfully isolated from bone essence of mice, the q-PCR results showed an increasing expression level of miR-146-5p in the process of MSC adipogenic differentiation. Compared with the control group, MSC transfected with miR-146b-5p mimic could up-regulate the expression of miR-146b-5p (P<0.001), while miR-146b-5p inhibitor transfection could down-regulate the endogenous miR-146b-5p expression (P<0.01). After culture for 14 d, the result of Oil red staining showed that the miR-146b-5p inhibitor could inhibit adipogenic differentiation, while the miR-146b-5p mimic could promote the adipogenic differentiation of MSC. After induction for 14 d, compared with control, the PPARγ and C/EBPα in mimic group were higher expressed PPARγ and C/EBPα (P<0.01). Compared with induced group, the PPAPγ and C/EBPα were lower expressed in inhibitor group (P<0.05). The results of Western blot showed that the expression level of PPARγ was high in minic group, and it was low in inhibitor group.
miR-146b-5p is up-regulated in the process of MSC adipogenic differentiation, and it promotes the adipogenesis of MSC originated from mouse bone essence.
探讨微小RNA-146b-5p(miR-146b-5p)对小鼠骨髓源性间充质干细胞(MSC)成脂分化的影响。
从C57BL/6小鼠骨髓中分离MSC。采用q-PCR检测MSC成脂分化过程中miR-146b-5p的表达水平;通过油红染色分析miR-146b-5p模拟物或抑制剂在小鼠骨髓源性MSC成脂分化过程中的作用,培养14天后,采用q-PCR检测C/EBPα和PPARγ的表达;采用蛋白质免疫印迹法检测miR-146b-5p转染后PPARγ的蛋白水平。
成功从小鼠骨髓中分离出MSC,q-PCR结果显示,MSC成脂分化过程中miR-146-5p表达水平升高。与对照组相比,转染miR-146b-5p模拟物的MSC可上调miR-146b-5p的表达(P<0.001),而转染miR-146b-5p抑制剂可下调内源性miR-146b-5p的表达(P<0.01)。培养14天后,油红染色结果显示,miR-146b-5p抑制剂可抑制成脂分化,而miR-146b-5p模拟物可促进MSC的成脂分化。诱导14天后,与对照组相比,模拟物组PPARγ和C/EBPα表达上调(P<0.01)。与诱导组相比,抑制剂组PPAPγ和C/EBPα表达下调(P<0.05)。蛋白质免疫印迹法结果显示,模拟物组PPARγ表达水平高,抑制剂组表达水平低。
miR-146b-5p在MSC成脂分化过程中表达上调,促进小鼠骨髓源性MSC的脂肪生成。