紫色悬钩子提取物对3T3-L1前脂肪细胞分化的影响

Effect of Rubus irritans Focke extract on differentiation of 3T3-L1 preadipocytes

目的研究紫色悬钩子提取物对3T3-L1前脂肪细胞成脂分化的影响,进一步探讨紫色悬钩子对脂肪细胞代谢的潜在机制,为肥胖症的预防和治疗提供新依据。方法MCI中压色谱柱分离制备紫色悬钩子提取物。MTT法检测细胞增殖,油红O染色及试剂盒检测分析细胞的脂质积累水平。Western blot检测过氧化物酶体增殖物激活受体γ(PPARγ)、CCAAT增强子结合蛋白(C/EBPα)、腺苷5′-单磷酸活化蛋白激酶(AMPK)及磷酸化的AMPK(p-AMPK)蛋白的表达。结果当紫色悬钩子提取物的质量浓度≤100 mg·L^(-1)时,对细胞活力的影响无明显变化(P<0.05),其中Fr.2、Fr.3、Fr.4、Fr.5可降低细胞中脂质的积累(P<0.05)。Fr.1低剂量组(10 mg·L^(-1))和Fr.4的高剂量组(100 mg·L^(-1))可以减少细胞中总胆固醇(TC)和三酰甘油(TG)含量,降低PPARγ、C/EBPα,升高p-AMPK的蛋白表达(P<0.05)。结论紫色悬钩子提取物可能是通过激活AMPK蛋白,抑制PPARγ信号通路相关蛋白的表达,从而降低细胞中脂质的积累。

Aim To study the effect of Rubus irritans Focke extract on adipogenic differentiation of 3T3-L1 preadipocytes,and to further explore the potential mechanism of Rubus irritans Focke on adipocyte metabolism,so as to provide a new basis for the prevention and treatment of obesity.Methods Rubus irritans Focke extract was separated and prepared by MCI medium pressure chromatographic column.MTT method was used to detect cell proliferation,and oil red O staining and kit test was used to to detect the level of lipid accumulation.Western blot was employed to detect the expressions of peroxisome proliferator-activated receptorγ(PPARγ),CCAAT enhancer binding proteinα(C/EBPα),adenosine 5′-monophosphate activated protein kinase(AMPK)and phosphorylated AMPK(p-AMPK)protein.Results When the concentration of Rubus irritans Focke extract was less than 100 mg·L^(-1),there was no significant effect on cell viability(P>0.05),but Fr.2,Fr.3,Fr.4 and Fr.5 could reduce the accumulation of lipids in cells(P<0.05).Fr.1 low dose group(10 mg·L^(-1))and Fr.4 high dose group(100 mg·L^(-1))could reduce the contents of total cholesterol(TC)and triglyceride(TG),decrease PPARγand C/EBPα,and increase the protein expression of p-AMPK(P<0.05).Conclusions Rubus irritans Focke contains active components that can reduce lipid production in adipocytes.The mechanism may be that the expression of proteins related to PPARγsignaling pathway is inhibited by activating AMPK protein,thus reducing lipid accumulation in adipocytes.