食品科学

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叶瓜参长链碱抑制3T3-L1前脂肪细胞分化作用机制

毛 磊,徐 慧,田迎樱,徐 杰,王玉明,王静凤*,薛长湖   

  1. 中国海洋大学食品科学与工程学院,山东 青岛 266003
  • 出版日期:2015-12-15 发布日期:2015-12-24

Suppressive Effect of Long-Chain Base from Sea Cucumber Cucumaria frondosa on the Differentiation of 3T3-L1 Preadipocytes and Underlying Mechanism

MAO Lei, XU Hui, TIAN Yingying, XU Jie, WANG Yuming, WANG Jingfeng*, XUE Changhu   

  1. College of Food Science and Engineering, Ocean University of China, Qingdao 266003, China
  • Online:2015-12-15 Published:2015-12-24

摘要:

目的:研究叶瓜参长链碱(long-chain base from the sea cucumber Cucumaria frondosa,Cf-LCB)对3T3-L1前脂肪细胞分化的作用,并探讨其作用机制。方法:以四甲基偶氮唑蓝(methyl thiazolyl tetrazolium,MTT)法检测Cf-LCB对3T3-L1前脂肪细胞增殖活性的影响;采用传统鸡尾酒法诱导3T3-L1前脂肪细胞分化为成熟脂肪细胞,分别采用油红O染色和甘油三酯(triglycerides,TG)含量测定法评价其对3T3-L1前脂肪细胞分化的影响;反转录实时荧光定量聚合酶链式反应(quantity real-time reverse transcript polymerase chain reaction,qRT-PCR)法检测脂肪细胞分化关键基因CCAAT增强子结合蛋白α(CCAAT/enhancer binding protein alpha,C/EBPα)、过氧化物酶体增殖物激活受体γ(peroxisome proliferators-activated receptors gamma,PPARγ)以及WNT/β-catenin通路关键基因WNT10b(wingless-type MMTV integration site family members)、卷曲蛋白1(frizzled protein1,FZ1)、低密度脂蛋白受体相关蛋白6(LDL-receptor-related protein6,LRP6)和β-连环蛋白(β-catenin)的mRNA表达水平;Western blotting法检测WNT/β-catenin通路关键基因LRP6和β-catenin的蛋白表达量。结果:Cf-LCB能显著抑制3T3-L1前脂肪细胞的增殖;抑制3T3-L1细胞脂滴形成以及C/EBPα和PPARγ mRNA表达;显著上调WNT/β-catenin通路关键基因FZ1、LRP6和β-catenin mRNA表达,对WNT10b的表达无影响;显著促进LRP6和β-catenin的蛋白表达,提高核内β-catenin含量。结论:Cf-LCB能够显著抑制3T3-L1前脂肪细胞分化,其作用机制与激活WNT/β-catenin通路有关。

关键词: 叶瓜参, 长链碱, 3T3-L1前脂肪细胞, 分化, WNT/&beta, -catenin信号通路

Abstract:

The aim of the present study was to observe the effect of Cf-LCB on the proliferation and differentiation of
3T3-L1 preadipocytes and to investigate the possible underlying mechanism. The effect of Cf-LCB on the proliferation
of 3T3-L1 preadipocytes was evaluated by MTT assay. After using differentiation cocktail to differentiate 3T3-L1
preadipocytes, the effect of Cf-LCB on the differentiation of 3T3-L1 preadipocytes was evaluated by oil red O-staining
and determining TG content. mRNA expression levels of key genes associated with differentiation and WNT/β-catenin
signal pathway in 3T3-L1 preadipocytes were measured by qRT-PCR. Expression levels of key proteins in WNT/β-catenin
signal pathway were measured by Western blotting. Results showed that Cf-LCB significantly inhibited the proliferation of
3T3-L1 preadipocytes, prevented the formation of lipid droplets in mature 3T3-L1 adipocytes and down-regulated C/EBPα
and PPARγ. The mRNA expression of FZ1, LRP6 and β-catenin was distinctly enhanced by Cf-LCB, suggesting activation
of WNT/β-catenin pathway. The increased protein expressions of key factors in WNT/β-catenin pathway including LRP6,
total and nuclear β-catenin further demonstrated that Cf-LCB could markedly suppress the proliferation and differentiation of
3T3-L1 preadipocytes by activating WNT/β-catenin pathway.

Key words: Cucumaria frondosa, long-chain base, 3T3-L1 preadipocytes, adipogenesis, WNT/β-catenin signal pathway

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