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6-姜酚通过激活3T3-L1脂肪细胞中的Wnt/β-连环蛋白信号通路对脂肪生成的抑制作用。

Inhibitory effect 6-gingerol on adipogenesis through activation of the Wnt/β-catenin signaling pathway in 3T3-L1 adipocytes.

作者信息

Li Chunbo, Zhou Lin

机构信息

Department of Gynaecology and Obstetrics, Shanghai First Maternity and Infant Hospital, Tongji University School of Medicine, Shanghai 200040, China.

Department of urology, Zhongshan Hospital, Fudan University, Shanghai 200032, China.

出版信息

Toxicol In Vitro. 2015 Dec 25;30(1 Pt B):394-401. doi: 10.1016/j.tiv.2015.09.023. Epub 2015 Oct 21.

Abstract

6-Gingerol has been reported to inhibit adipogenesis and lipid content accumulation. However, the mechanism of its anti-adipogenic effect remains unclear. Our aim is to investigate the molecular mechanism of the anti-adipogenic effect of 6-gingerol. The lipid content in adipocytes was measured by Oil Red O staining and cell viability was analyzed by MTT assay. The extent of suppression of differentiation by 6-gingerol was characterized by measuring the triglyceride content and GPDH activity. The regulation of adipogenic markers and the components of the Wnt/β-catenin pathway were analyzed by real-time PCR and Western blotting. The nuclear location of β-catenin was identified using immunofluorescence assay. Small interfering RNA transfection was conducted to elucidate the crucial role of β-catenin in anti-adipogenic effect of 6-gingerol. Our results showed that 6-gingerol inhibited the adipogenesis and lowered the mRNA expression levels of transcription factors and the key lipogenic enzymes in 3T3-L1 cells. The effect of 6-gingerol on adipogenic differentiation was accompanied by stimulating the activation of the Wnt/β-catenin signaling. In addition, we found that 6-gingerol induced phosphorylations of glycogen synthase kinase-3β(GSK-3β), and promoted the nuclear accumulation of β-catenin. Importantly, the inhibitory effect of 6-gingerol on adipogenic differentiation was reversed after the siRNA knockdown of β-catenin was added. Our findings demonstrated that 6-gingerol inhibits the adipogenic differentiation of 3T3-L1 cells through activating the Wnt/β-catenin signaling pathway.

摘要

据报道,6-姜酚可抑制脂肪生成和脂质含量积累。然而,其抗脂肪生成作用的机制仍不清楚。我们的目的是研究6-姜酚抗脂肪生成作用的分子机制。通过油红O染色测量脂肪细胞中的脂质含量,并通过MTT法分析细胞活力。通过测量甘油三酯含量和甘油磷酸脱氢酶(GPDH)活性来表征6-姜酚对分化的抑制程度。通过实时PCR和蛋白质印迹分析脂肪生成标志物和Wnt/β-连环蛋白信号通路的组成成分的调控情况。使用免疫荧光测定法确定β-连环蛋白的核定位。进行小干扰RNA转染以阐明β-连环蛋白在6-姜酚抗脂肪生成作用中的关键作用。我们的结果表明,6-姜酚抑制3T3-L1细胞的脂肪生成,并降低转录因子和关键脂肪生成酶的mRNA表达水平。6-姜酚对脂肪生成分化的影响伴随着刺激Wnt/β-连环蛋白信号的激活。此外,我们发现6-姜酚诱导糖原合酶激酶-3β(GSK-3β)的磷酸化,并促进β-连环蛋白的核积累。重要的是,添加β-连环蛋白的小干扰RNA敲低后,6-姜酚对脂肪生成分化的抑制作用被逆转。我们的研究结果表明,6-姜酚通过激活Wnt/β-连环蛋白信号通路抑制3T3-L1细胞的脂肪生成分化。

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