天津医药

天津医药 ›› 2015, Vol. 43 ›› Issue (12): 1345-1348.doi: 10.11958/j.issn.0253-9896.2015.12.001

• 细胞与分子生物学 •    下一篇

川续断皂苷Ⅵ对小鼠骨髓基质细胞ST-2 成脂分化的影响及其机制研究

王海晓1，崔壮2，王宝利3，边育红1，郑纺1△   

  1. 1天津中医药大学中西医结合学院（邮编300193）；2天津医科大学公共卫生学院；3天津医科大学代谢病医院内分泌研究所
  • 收稿日期:2015-07-06 修回日期:2015-08-28 出版日期:2015-12-15 发布日期:2015-12-11
  • 通讯作者: 郑纺 E-mail:zhengfang_1979@163.com
  • 作者简介:王海晓（1989），男，硕士在读，主要从事中西医结合防治代谢性骨病方面的研究
  • 基金资助:
    国家自然科学基金资助项目（81102543,81200611）；中国博士后基金面上项目（2012M520588）

Effect of Asperosaponin Ⅵ on adipocyte differentiation in ST-2 cells and its underlying mechanisms

WANG Haixiao1，CUI Zhuang2，WANG Baoli3，BIAN Yuhong1，ZHENG Fang1△   

  1. 1 College of Integrated Traditional and Western Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China; 2 College of Public Health，Tianjin Medical University; 3 Key Laboratory of Hormone and Development（Ministry of Health），Metabolic Diseases Hospital & Institute of Endocrinology, Tianjin Medical University
  • Received:2015-07-06 Revised:2015-08-28 Published:2015-12-15 Online:2015-12-11
  • Contact: ZHENG Fang E-mail:zhengfang_1979@163.com

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摘要： 目的观察川续断皂苷Ⅵ（Asperosaponin Ⅵ，ASAⅥ）对脂肪细胞分化的影响及Wnt 通路的调节。方法以小鼠骨髓基质细胞系ST-2 为研究对象，将其分为对照组、成脂分化组以及4 个不同剂量的ASAⅥ组。其中对照组加入溶媒，成脂分化组加入成脂诱导分化试剂，ASAⅥ组除加入成脂诱导分化试剂外，使用不同浓度（10-7、10-6、10-5、 10-4 mol/L）的ASAⅥ干预细胞。各组细胞处理5 d 后，行油红O 染色，观察脂滴形成情况并计算成脂率进行定量分析；荧光定量PCR（FQ-PCR）检测成脂相关基因PPARγ、FABP4 和Wnt/β-连环素（β-catenin）通路蛋白β-catenin 的 mRNA 表达水平；Wnt 通路抑制剂DKK1 干预诱导成脂分化的ST-2 细胞5 d，FQ-PCR 检测ASAⅥ所调节的 PPARγ、FABP4 和β-catenin mRNA 表达水平。结果与成脂分化组相比，10-5 mol/L 和10-4 mol/L ASAⅥ组中分化的脂肪细胞显著减少，10-5、10-4 mol/L ASAⅥ明显抑制PPARγ、FABP4 的mRNA 表达，但上调β-catenin mRNA 表达。 DKK1 能够逆转ASAⅥ对ST-2 细胞成脂分化的抑制作用，促进PPARγ、FABP4 的mRNA 表达，抑制β-catenin 的 mRNA 表达。结论ASAⅥ能显著抑制ST-2 细胞的成脂分化，这一作用可能是通过Wnt/β-catenin 通路的激活所介导的。

关键词: 成脂分化, β连环素, 川续断皂苷Ⅵ, 骨髓基质细胞, Wnt 信号通路

Abstract: Objective The effect of Asperosaponin Ⅵ（ASAⅥ）on adipocyte differentiation and the involvement of Wnt signal pathway was investigated. Methods The murine bone marrow stromal cell line ST- 2 were divided into 6 groups: control group, adipocyte differentiation group, and 4 different doses of ASAⅥ groups. Control group was exposed to the vehicle, adipocyte differentiation group was exposed to adipogenic reagent, and those 4 ASAⅥ groups were treated with different concentration（10-7, 10-6, 10-5, 10-4 mol/L）of ASAⅥ after adipocyte differentiation induction. 5 days later, oil red O staining was performed to calculate adipocyte rate. Then mRNA transcription levels of PPARγ, FABP4 genes and β-catenin that were Wnt/β-catenin signaling pathway proteins were examined by FQ-PCR. Then Wnt pathway inhibitor DKK1 was supplemented into ST-2 cells treated with 10-4 mol/L ASAⅥ for 5 days. After that FQ-PCR was used to detect whether tran⁃ scription levels of PPARγ, FABP4 and β-catenin in ST-2 cells were changed. Results Compared with adipocyte differenti⁃ ation group 10-5 mol/L and 10-4 mol/L ASAⅥ treatments greatly down-regulated the number of lipid droplets and markedly inhibited transcription levels of adipocyte characterization transcription factors included PPARγ, FABP4 while up-regulat⁃ ed transcription level of β-catenin in ST-2 cells. DKK1 can reverse the inhibitory effect of ASAⅥ on adipocyte differentia⁃ tion in ST-2 adipocyte. The transcription levels of PPARγ and FABP4 were up-regulated significantly while transcription level of β-catenin was inhibited. Conclusion ASAⅥ blocks adipocyte differentiation in ST-2 cells which might be medi⁃ ated through activating Wnt/β -catenin signaling pathway.

Key words: adipocyte differentitation, beta catenin, Asperosaponin Ⅵ, bone marrow stromal cells, Wnt signal pathway