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葛根对3T3-L1脂肪细胞的葡萄糖和脂质代谢进行调节以改善胰岛素抵抗

[Kudzu root (Ge-Gen) regulates on glucose and lipid metabolism to ameliorating insulin resistance on 3T3-L1 adipocytes].

作者信息

Luo Xin-Xin, Xu Guo-Liang, Li Yu, Li Bing-Tao, Tu Jun

机构信息

Jiangxi Province Key Laboratory of Traditional Chinese Medicine Etiopathogenisis, Research Center for Differentiation and Development of Traditional Chinese Medicine Basic Theory, Jiangxi University of Traditional Chinese Medicine, Nanchang 330004, China.

出版信息

Zhongguo Zhong Yao Za Zhi. 2016 Jul;41(14):2687-2694. doi: 10.4268/cjcmm20161420.

Abstract

This study aimed to explore the mechanism of Chinese traditional medicine, Kudzu root(Chinese name:Ge-Gen; Latin name: Puerariae Lobatae Radix) how to improving insulin resistance (IR) through the regulation of the glucose and lipid metabolism in the IR-3T3-L1 adipocytes. After the 3T3-L1 mouse preadipocytes were differentiated into mature adipocytes, IR model(IR-3T3-L1) was built with 1 μmol•L-1 dexamethasone treatment for 96 h. IR adipocytes were treated with different concentrations (5%,10% and 15%) of Ge-Gen containing serum (GG-CS)for 12 h or 24 h, whereas rosiglitazone group as positive control in this study. The glucose contents in cell culture supernatants were detected by glucose oxidase assay and the intracellular triglyceride (TG) contents were measured by glycerol phosphate oxidase assay respectively.The mRNA expression levels of PPARγ, ADPN, GLUT4, LPL, FABP4 and FASn gene were determined by real-time quantitative PCR(qPCR).Results showed that IR-3T3-L1 adipocytes significantly increased glucose consumption (P<0.01)and decreased TG contents (P<0.01) as compared with the normal control group, the glucose consumption significantly increased with the treatment of GG-CS (P<0.01) by dose-dependent and time-dependent manners,whereas the intracellular TG content was sigificantly decreased (P<0.01) by dose-dependent manner.qPCR analysis revealed that 10% and 15% GG-CS significantly up-regulated the mRNA expression level of PPARγ, ADPN and GLUT4 (P<0.01) with the same dose-dependent manner,whereas the GLUT4 mRNA expression was showed similar expression pattern with the treatment of 10% and 15% GG-CS (P<0.01).We also detected the mRNA expression levels of several important lipid-metabolizing enzymes such as LPL, FASn and FABP4 by PPARγ regulation. 15% GG-CS elevated LPL mRNA expression (P<0.05);10% and 15% GG-CS enhanced the FASn mRNA expression (P<0.01), whereas 5%,10% and 15% GG-CS down-regulated FABP4 mRNA expression (P<0.01). Together, our results indicated that GG could regulate the glucose and lipid metabolism to ameliorate IR with multi-target manners in 3T3-L1 adipocytes.

摘要

本研究旨在探讨中药葛根(中文名:葛根;拉丁名:Puerariae Lobatae Radix)如何通过调节胰岛素抵抗(IR)-3T3-L1脂肪细胞中的糖脂代谢来改善胰岛素抵抗(IR)。将3T3-L1小鼠前脂肪细胞分化为成熟脂肪细胞后,用1μmol•L-1地塞米松处理96小时建立IR模型(IR-3T3-L1)。用不同浓度(5%、10%和15%)的含葛根血清(GG-CS)处理IR脂肪细胞12小时或24小时,而罗格列酮组作为本研究的阳性对照。分别用葡萄糖氧化酶法检测细胞培养上清液中的葡萄糖含量,用甘油磷酸氧化酶法检测细胞内甘油三酯(TG)含量。通过实时定量PCR(qPCR)测定PPARγ、ADPN、GLUT4、LPL、FABP4和FASn基因的mRNA表达水平。结果显示,与正常对照组相比,IR-3T3-L1脂肪细胞的葡萄糖消耗量显著增加(P<0.01),TG含量显著降低(P<0.01);GG-CS处理后,葡萄糖消耗量呈剂量和时间依赖性显著增加(P<0.01),而细胞内TG含量呈剂量依赖性显著降低(P<0.01)。qPCR分析显示,10%和15%的GG-CS以相同的剂量依赖性方式显著上调PPARγ、ADPN和GLUT4的mRNA表达水平(P<0.01),而GLUT4 mRNA表达在10%和15%的GG-CS处理下呈现相似的表达模式(P<0.01)。我们还通过PPARγ调节检测了几种重要的脂质代谢酶如LPL、FASn和FABP4的mRNA表达水平。15%的GG-CS提高了LPL mRNA表达(P<0.05);10%和15%的GG-CS增强了FASn mRNA表达(P<0.01),而5%、10%和15%的GG-CS下调了FABP4 mRNA表达(P<0.01)。总之,我们的结果表明,GG可以通过多靶点方式调节3T3-L1脂肪细胞中的糖脂代谢以改善IR。

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