染料木苷对3T3-L1脂肪细胞分化以及AMPK磷酸化的影响

Impacts of Genistin on 3T3-L1 Adipocyte Differentiation and AMPK Phosphorylation

目的观察染料木苷(Genistin,GIN)对3T3-L1前脂肪细胞细胞分化以及分化过程中腺苷酸活化蛋白激酶(AMP-activated protein kinase,AMPK)磷酸化程度的影响。方法用激素鸡尾酒法诱导3T3-L1前脂肪细胞分化,用MTT法观察GIN对细胞生存率的影响,以油红O染色法镜下观察3T3-L1前脂肪细胞分化的情况,并通过比色分析法检测GIN对细胞内脂滴堆积量的影响;用Western blotting方法分别检测对照组和给药组细胞分化过程中细胞中AMPK,P-AMPK(Thr172)蛋白表达量,借以计算AMPK磷酸化比率。结果分化诱导后,对照组脂肪细胞呈典型的成熟脂肪细胞表型,形成"戒指环"结构,经100μM GIN干预后,脂滴数量减少,体积减小,通过比色分析法量化后显示100μM GIN可减少胞内脂质的堆积(P<0.05)。与对照组相比,给药组从第4天开始上调p-AMPK(Thr172)/AMPK比率,并持续到第10天(P<0.05)。结论 GIN可通过激活AMPK抑制3T3-L1脂肪细胞分化。

Objective To observe the impacts of genistin （GIN）on 3T3 -L1 preadipocyte differentiation and AMPK phosphorylation during differentiation. Methods The hormone cocktail method was adopted to induce 3T3 - L1 preadipocyte differentiation. MTT method was used to observe the impacts of GIN on cell survival rate. The oil red O staining method was applied for the microscopic observation of 3T3 - L1 preadipo- cyte differentiation. Colorimetric analysis method was used to detect the impacts of GIN on intracellular lipid droplets deposition. Western blotting method was used to detect the expressions of AMPK and P - AMPK （Thr172） during the cellular differentiation in the control group and medication group so as to calculate the proportion of AMPK phosphorylation. Results After differentiation and induction, in the control group, the adipocytes displayed the typical mature adipocyte phenotype ,in the structure as＂ ring＂. After 100 μM GIN intervention, the droplets amount and volume were reduced. The quantification of colorimetric analysis indicated that 100 μM GIN reduced intracellular lipid accumulation （ P 〈 0.05 ）. Compared with the control group, since the 4th day in the medication group, the ratio of P - AMPK（ Thr172 ） and AMPK was up - regulated till the 10th day （ P 〈 0.05 ）. Conclusion GIN inhibits 3T3 - L1 adipocyte differentiation via activating AMPK.