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、50μmol/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,10μmol/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。
In order to study the effect of puerarin on differentiation of 3 T3-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 50μmol/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 3 T3-L1 adipocytes,respectively.The mRNA expression of CCAAT-enhancer binding proteinα（C/EBPα）and peroxisome proliferator-activated receptorγ（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 10μmol/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/EBPαand PPARγ（P0.01）.Further studies showed that stimulation of puerarin enhanced the phosphorylation level of the Akt signaling pathwaySer473 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 3 T3-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.
出处
《中国畜牧兽医》
CAS
北大核心
2018年第1期57-64,共8页
China Animal Husbandry & Veterinary Medicine
基金
吉林省农业科学院创新工程项目(CXGC2017JQ001)
