葛根素对Akt通路调控3T3-L1脂肪细胞分化的影响

doi:10.16431/j.cnki.1671-7236.2018.01.007

摘要/Abstract

摘要：

为了研究葛根素（puerarin）对3T3-L1前体脂肪细胞分化的影响，探索其在成脂分化过程中的潜在作用机制，试验在脂肪形成过程中将0、10、50 μmol/L葛根素加入到诱导分化培养基中诱导分化，分别通过油红O染色法及甘油三酯酶法检测葛根素对3T3-L1脂肪细胞的脂滴积累、甘油三酯的影响；采用实时荧光定量PCR检测脂肪细胞中CCAAT-增强子结合蛋白α（C/EBPα）和过氧化物酶体增殖物激活受体γ（PPARγ）的mRNA表达量，Western blotting检测脂肪形成相关转录因子及Akt信号通路的蛋白水平的表达量。结果表明，10 μmol/L葛根素极显著增加了成熟脂肪细胞中脂滴和甘油三酯（TG）的积聚，极显著促进了脂肪形成相关转录因子C/EBPα和PPARγ的mRNA和蛋白水平的表达量（P<0.01）。进一步研究发现，与对照组相比，葛根素的刺激可增强Akt信号通路Ser473蛋白的磷酸化表达水平，表明葛根素对成脂分化过程的促进作用很大程度上是通过Akt信号通路的磷酸化来实现的。综上所述，葛根素能够促进3T3-L1前体脂肪细胞的分化，改善胰岛素敏感性，其作用机制与激活Akt信号通路Ser473位点的磷酸化水平有关。本试验结果可为研究胰岛素的效应机制提供新见解，为胰岛素抵抗相关疾病的治疗提供新思路。

关键词: 葛根素; 脂肪生成; 3T3-L1; Akt信号通路

Abstract:

In order to study the effect of puerarin on differentiation of 3T3-L1 pre-adipocytes, explore the potential mechanism of puerarin in adipogenic differentiation. Puerarin was added into the medium to induce differentiation with final concentration of 0,10 and 50 μmol/L in the process of adipogenesis. The effects of puerarin on accumulation of lipid droplets and the content of triglyceride were measured by oil red O staining and triglyceride assay kit in 3T3-L1 adipocytes, respectively. The mRNA expression of CCAAT-enhancer binding protein α (C/EBPα) and peroxisome proliferator-activated receptor γ (PPARγ) in adipocytes were detected by quantitative Real-time PCR, and the protein expression levels of adipogenic transcription factor and Akt signaling pathway were qualified by Western blotting. The results showed that 10 μmol/L puerarin extremely significantly increased the accumulation of lipid droplets and triacylglycerol (TG) in mature adipocytes, which extremely significantly promoted the mRNA and protein expression levels of adipogenic transcription factor C/EBPα and PPARγ (P<0.01). Further studies showed that stimulation of puerarin enhanced the phosphorylation level of the Akt signaling pathway Ser473 protein compared to control group, suggested that the phosphorylation of Akt signaling pathway played a major role in promoting adipogenic differentiation. In conclusion, puerarin could promote the differentiation of 3T3-L1 pre-adipocytes and improve the sensitivity of insulin, and its mechanism was related to the phosphorylation level of Ser473, which activated Akt signaling pathway. This research would provide a new insights into the mechanism of insulin and a new ideas for treatment of insulin resistance related diseases.

Key words: puerarin; adipogenesis; 3T3-L1; Akt pathway

中图分类号:

Q813.1⁺1

云巾宴, 于永生, 曹阳, 张立春, 金海国, 罗晓彤, 魏天, 李欣, 金鑫. 葛根素对Akt通路调控3T3-L1脂肪细胞分化的影响[J]. 《中国畜牧兽医》, 2018, 45(1): 57-64.

YUN Jinyan, YU Yongsheng, CAO Yang, ZHANG Lichun, JIN Haiguo, LUO Xiaotong, WEI Tian, LI Xin, JIN Xin. Effect of Puerarin on Regulation Adipocytes Differentiation Through Akt Pathway in 3T3-L1 Cells[J]. , 2018, 45(1): 57-64.

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