广藿香水提液对3T3-L1前脂肪细胞增殖、分化和改善胰岛素抵抗的作用

Effect of Pogostemon cablin extract on proliferation and differentiation of 3T3-L1 preadipocytes and improvement of insulin resistance

本研究旨在探讨藿香(Pogostemon cablin,PC)水提取物在促进脂肪细胞分化和改善脂肪细胞胰岛素抵抗(IR)中的作用。于诱导分化第1天,在3T3-L1前脂肪细胞培养板中加入不同质量浓度PC水提液(0.5,1.0,2.5,5.0 g/L),第8天,采用油红O染色和染色比色法对3T3-L1脂肪细胞的脂质沉积进行观察和分析,通过ELISA检测细胞内甘油三酯(triglyceride,TG)水平,Western blot测定过氧化物酶体增殖物激活的受体γ-2(peroxisome proliferator-activated receptor-γ2,PPARγ2)蛋白表达情况,通过四甲基偶氮唑盐(MTT)法分析PC对3T3-L1前脂肪细胞增殖和分化的影响,葡萄糖氧化酶-过氧化物酶法检测分化成熟的脂肪细胞的葡萄糖消耗情况。结果表明,PC水提液能抑制3T3-L1脂肪细胞的增殖(P<0.01),并促进细胞向成熟脂肪细胞分化(P<0.05)。随着PC质量浓度的增加,3T3-L1前脂肪细胞内脂质沉积逐渐增加,TG水平逐渐上升(P<0.05)。与诱导分化组比较,PC水提液组(1.0,2.5 g/L)PPARγ2蛋白表达显著上升(P<0.05)。与成熟3T3-L1脂肪细胞IR组对比发现,PC能极显著促进成熟脂肪细胞的葡萄糖消耗(P<0.01),改善IR。结果表明,PC可抑制3T3-L1前脂肪细胞的增殖并促进其分化,促进细胞内脂滴的积累并调节糖脂代谢,从而改善IR。

The aim of this study is to explore the effect of Pogostemon cablin(PC)on the promotion of 3 T3-L1 preadipocytes differentiation and the regulation of glucose and lipid metabolism in adipocytes.Different concentrations of PC(0.5,1.0,2.5 and 5.0 g/L)were added into cell culture medium on the first day.On the eighth day,lipid droplets accumulated in cytoplasm of the differentiated preadipocytes were observed by oil red O staining and quantified by colorimetry.ELISA method was used to detect intracellular triglyceride(TG)levels.Western blot was used to determine the protein expression of PPARγ2.MTT assay was used to analyze the effect of PC on the proliferation and differentiation of 3 T3-L1 cells.Glucose oxidase-peroxidase method was used to detect glucose concentration in mature adipocytes.The results showed that PC could inhibit the proliferation of 3 T3-L1 preadipocytes(P<0.01)and promote the differentiation of cells to mature adipocytes(P<0.05).With the increase of PC concentration,lipid storage gradually increased and TG level gradually increased(P<0.05).Compared with the induced differentiation group,the expression of PPARγ2 protein in PC group(1.0 and 2.5 g/L)was significantly increased(P<0.05).Compared with the insulin-resistant mature 3 T3-L1 adipocyte group,it was found that PC could significantly promote glucose consumption and improve insulin resistance in mature adipocytes(P<0.01).The results suggest that PC can promote the differentiation of 3 T3-L1 preadipocytes,promote the accumulation of intracellular lipid droplets and regulate the metabolism of glucose and lipids,thereby improving insulin resistance.