Spry1对脂肪细胞分化的调控作用

目的探讨利用siRNA下调骨髓基质细胞系ST2细胞中Spry1的内源性表达对脂肪细胞分化的调控作用。方法设计Spry1的靶向siRNA作为实验组，以转染Control siRNA作为对照组。在ST2细胞中转染Spry1 siRNA和Control siRNA并进行成脂诱导，应用qRT-PCR检测2组细胞中的Spry1和成脂特异性因子过氧化物酶体增殖物激活受体（PPAR）γ、CCAAT增强子结合蛋白（C/EBP）α、脂肪细胞表征因子FABP4、脂肪因子adipsin的mRNA表达水平。脂肪细胞分化成熟后，进行油红O染色，显微镜下观察脂肪细胞的染色情况及Spry1siRNA对ST2细胞成脂分化的影...

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Bibliographic Details
Published in天津医药 Vol. 45; no. 11; pp. 1171 - 1174
Main Author 岑运珠;刘颖;杨威利;王宝利
Format Journal Article
LanguageChinese
Published 天津医科大学代谢病医院内分泌研究所,卫生部激素与发育重点实验室 300070 2017
天津医科大学口腔医院牙体牙髓科%天津医科大学口腔医院牙体牙髓科%天津医科大学基础医学院%天津医科大学代谢病医院内分泌研究所,卫生部激素与发育重点实验室,300070
Subjects
间质干细胞;成脂分化;脂细胞;RNA,小分子干扰;Spry1
成脂分化
RNA,小分子干扰
adipogenesis
RNA
mesenchymal stem cells
small interfering
Spry1
adipocytes
脂细胞
间质干细胞
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ISSN0253-9896
DOI10.11958/20170715

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Summary:目的探讨利用siRNA下调骨髓基质细胞系ST2细胞中Spry1的内源性表达对脂肪细胞分化的调控作用。方法设计Spry1的靶向siRNA作为实验组，以转染Control siRNA作为对照组。在ST2细胞中转染Spry1 siRNA和Control siRNA并进行成脂诱导，应用qRT-PCR检测2组细胞中的Spry1和成脂特异性因子过氧化物酶体增殖物激活受体（PPAR）γ、CCAAT增强子结合蛋白（C/EBP）α、脂肪细胞表征因子FABP4、脂肪因子adipsin的mRNA表达水平。脂肪细胞分化成熟后，进行油红O染色，显微镜下观察脂肪细胞的染色情况及Spry1siRNA对ST2细胞成脂分化的影响。运用异丙醇萃取染色的脂肪细胞中的油红O并测定波长在520nm处的光密度（OD）值。结果转染Spry1siRNA于ST2细胞后，与转染ControlsiRNA的对照组相比，Spry1基因的表达水平明显下调，Spry1siRNA可抑制脂肪细胞的分化，油红O染色显示实验组的脂肪细胞明显减少且OD值低于对照组；转染Spry1siRNA实验组的成脂特异性因子PPARγ、C/EBPα、FABP4、adipsin的mRNA表达水平显著下调，差异有统计学意义（P<0.05）。结论Spry1siRNA可有效抑制前体细胞向脂肪细胞分化。
Bibliography:mesenchymal stem cells; adipogenesis; adipocytes; RNA, small interfering; Spry1
Objective To investigate the effect of Spry1on adipocyte differentiation from ST2cells by using siRNA.Methods Synthesized siRNA targeting Spry1was used as experimental group,and control siRNA was used as control group.Spry1siRNA and control siRNA were transfected into ST2cells,then treating with adipogenic medium to induce adipocyte differentiation.The mRNA expression levels of Spry1and adipocyte differentiation-specific genes PPARγ(peroxisome proliferator-activated receptor gamma),C/EBPα(CCAAT enhancer binding proteinα),FABP4(adipocyte marker gene fatty acid binding protein4)and adipsin were examined by quantitative real-time PCR.The mature adipocytes were stained with oil red O,the staining adipocytes were observed by microscope,then understanding the effect of Spry1siRNA on adipocyte differentiation.In addition,oil red O of the staining adipocytes was extracted with isopropanol,optical density(OD)values of oil red O were measure
ISSN:0253-9896
DOI:10.11958/20170715