解剖学报

解剖学报 ›› 2017, Vol. 48 ›› Issue (4): 416-420.doi: 10.16098/j.issn.0529-1356.2017.04.008

• 细胞和分子生物学 • 上一篇    下一篇

Krüppel样因子4在猪脂肪间充质干细胞成脂分化过程中的表达模式

李秀 李方正 郇延军 高霞 姜忠玲 宋学雄*   

  1. 青岛农业大学动物科技学院，山东 青岛 266109
  • 收稿日期:2016-11-16 修回日期:2017-02-06 出版日期:2017-08-06 发布日期:2017-08-06
  • 通讯作者: 宋学雄 E-mail:xuexiongs@yahoo.com.cn
  • 基金资助:
    姜黄素调控猪间充质干细胞向脂肪细胞分化及Krüppel样因子转录网络的介导作用

Expression pattern of Krüppel-like factor 4 during adipogenic differentiation of porcine adipogenic mescenchymal stem cells

LI Xiu LI Fang-zheng HUAN Yan-jun GAO Xia JIANG Zhong-ling SONG Xue-xiong*   

  1. College of Animal Science and Technology, Qingdao Agricultural University, Shandong Qingdao 266109, China
  • Received:2016-11-16 Revised:2017-02-06 Online:2017-08-06 Published:2017-08-06
  • Contact: SONG Xue-xiong E-mail:xuexiongs@yahoo.com.cn

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摘要：

目的 探讨猪脂肪间充质干细胞（AMSCs）成脂分化过程中Krüppel样因子4（KLF4）的表达模式。方法 采用F3代以上猪AMSCs进行成脂诱导分化，用油红O染色提取法检测成脂分化程度，用RT-PCR和Real-time PCR检测KLF4的表达模式。 结果 猪AMSCs成脂诱导分化，诱导3 d开始出现脂肪滴，诱导 10 d时成脂分化率达60%；RT-PCR检测，在诱导分化2、4、8、12和16 d时都能检测出KLF4的表达；经Real-time PCR检测，上述各诱导分化时间点的表达量分别为对照组的1.6657±0.2269、2.6790±0.0524、2.6293±0.0280、2.3633±0.1568和2.6146±0.1575倍。这些结果表明，KLF4的表达在猪AMSCs成脂诱导分化的第2天就有显著上调，表达高峰出现在成脂分化的第4天，此后持续高表达。 结论 在猪AMSCs成脂分化过程中，从早期到晚期，KLF4发挥持续性的调控作用。

关键词: 脂肪间充质干细胞, 成脂分化, Krüppel样因子4, 实时定量聚合酶链反应, 猪

Abstract:

Objective To investigate the expression pattern of Krüppel-like factor 4 (KLF4) during adipogenic differentiation of porcine adipogenic mescenchymal stem cells (AMSCs). Methods More than F3 generation of porcine AMSCs were used during adipogenic differentiation, the level of adipogenic differentiation was detected by oil red O staining extraction assay, the expression of KLF4 was analyzed by RT-PCR and Real-time PCR. Results During the porcine AMSCs adipogenic differentiation, the lipid droplets began to appear on 3 days after induced, and the rate of adipogenic differentiation was about 60% on 10 days after induced;During the day of 2, 4, 8, 12 and 16 induced, the expression of KLF4 was detected in each detect time by RT-PCR; Quantitative analysis of the KLF4 expression levels on the above days by Real-time PCR, showed 1.6657±0.2269, 2.6790±0.0524, 2.6293±0.0280, 2.3633±0.1568 and 2.6146±0.1575 folds high compared with the uninduced control group. These results suggested that the expression of KLF4 was significantly increased on the day 2 induced, the highest expression was on the day 4 induced, and remained high expression after then. Conclusion During porcine AMSCs adipogenic differentiation, KLF4 may play a continuous regulatory role from the early to later period of induction.

Key words: Adipogenic mescenchymal stem cell, Adipogenic differentiation, Krüppel-like facter 4, Real-time PCR, Porcine