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小檗碱作用下3T3-L1细胞中miRNA-27a和miRNA-27b上调的功能研究

Functional study of the upregulation of miRNA-27a and miRNA-27b in 3T3-L1 cells in response to berberine.

作者信息

Wu Yue-Yue, Huang Xin-Mei, Liu Jun, Cha Ying, Chen Zao-Ping, Wang Fang, Xu Jiong, Sheng Li, Ding He-Yuang

机构信息

Department of Endocrinology, The Fifth People's Hospital of Shanghai Affiliated to Fudan University, Shanghai 200240, P.R. China.

出版信息

Mol Med Rep. 2016 Sep;14(3):2725-31. doi: 10.3892/mmr.2016.5545. Epub 2016 Jul 25.

Abstract

Berberine is the major active component of Rhizoma Coptidis derived from a traditional Chinese herbal medicine and is known to regulate micro (mi)RNA levels, although the mechanism for this action remains unknown. The present study confirmed that treatment of 3T3‑L1 cells with berberine inhibited cell viability and differentiation in a dose‑ and time‑dependent manner, and significantly increased the mRNA expression levels of miRNA‑27a and miRNA‑27b. In addition, in 3T3‑L1 cells treated with berberine, overexpression of miRNA‑27a and miRNA‑27b improved the berberine-mediated inhibition of cell differentiation and reduction of triglyceride contents. By contrast, miRNA‑27a and miRNA‑27b inhibitors attenuated the berberine‑mediated inhibition of cell differentiation and reduction of triglyceride contents. Additionally, peroxisome proliferator‑activated receptors (PPAR)‑γ was confirmed to be a target of miRNA‑27a in the 3T3‑L1 cells. A dual‑luciferase reporter assay indicated that the expression of PPAR‑γ was negatively regulated by miRNA-27a. These findings may provide novel mechanistic insight into the antiobesity effects of certain compounds in traditional Chinese herbal medicine.

摘要

黄连素是源自传统中草药黄连的主要活性成分,已知其可调节微小(mi)RNA水平,但其作用机制尚不清楚。本研究证实,用黄连素处理3T3-L1细胞会以剂量和时间依赖性方式抑制细胞活力和分化,并显著提高miRNA-27a和miRNA-27b的mRNA表达水平。此外,在用黄连素处理的3T3-L1细胞中,miRNA-27a和miRNA-27b的过表达改善了黄连素介导的细胞分化抑制和甘油三酯含量降低。相比之下,miRNA-27a和miRNA-27b抑制剂减弱了黄连素介导的细胞分化抑制和甘油三酯含量降低。此外,过氧化物酶体增殖物激活受体(PPAR)-γ被证实是3T3-L1细胞中miRNA-27a的靶标。双荧光素酶报告基因检测表明,PPAR-γ的表达受到miRNA-27a的负调控。这些发现可能为某些传统中草药化合物的抗肥胖作用提供新的机制见解。

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