杨梅素联合围脂滴蛋白对脂肪细胞脂解的影响

Effect of PLIN1 and myricetin on lipolysis of 3T3-L1 adipocytes

目的研究杨梅素(Myric)与围脂滴蛋白(PLIN1)联合作用对3T3-L1脂肪细胞脂解的影响。方法常规培养及诱导分化3T3-L1前脂肪细胞,筛选出Myric最佳干预浓度和时间。Myric联合sh-RNA干扰载体对已诱导分化成熟的3T3-L1脂肪细胞进行联合干预,实验共分为四组:联合干预组(Myric+sh-RNA)、转染组(sh-RNA)、杨梅素组(Myric)和空白组。采用油红O进行脂滴染色,观察脂滴形态;采用酶学方法检测细胞中三酰甘油(TG)和甘油含量,了解细胞脂解情况;采用Western blot检测脂肪细胞中PLIN1A、三酰甘油脂肪酶(ATGL)和激素敏感性脂肪酶(HSL)的表达量。结果以浓度为100μmol/L的Myric干预72 h时,细胞内TG含量最低,甘油含量最高,脂解效果最佳,与其他浓度和干预24 h比较,差异有统计学意义(P <0.05)。联合干预后,与Myric组和sh-RNA组比较,Myric+sh-RNA组细胞内TG含量下降,甘油含量升高,脂滴形态变小,数量减少,差异有统计学意义(P <0.05)。同时,Myric+sh-RNA组PLIN1A蛋白表达量降低,ATGL和HSL蛋白表达量升高,差异有统计学意义(P <0.05)。结论 Myric联合PLIN1可以更有效降低PLIN1表达量,提高ATGL与HSL的表达量,从而提高脂解效率。

Objective To investigate the effect of PLIN1 and myricetin on lipolysis of 3T3-L1 adipocytes and potential mechanisms.Methods 3T3-L1 preadipocytes were routinely cultured and differentiated into adipocytes.Mature 3T3-L1 adipocytes were treated with Myric under gradient concentration and time duration for optimal situation.Mature 3T3-L1 adipocytes were treated with Myric and/or sh-RNA of PLIN1 and were divided into four groups:combined treatment group(Myric+sh-RNA),transfection group(sh-RNA),myricetin group(Myric),and blank control group.Oil red O staining was performed to observe the morphology of lipid droplets.Enzymatic method was utilized to determine the contents of intracellular TG and glycerol.Western blot analysis was employed for identification of expression of PLIN1A,ATGL and HSL.Results Myric concentration at 100μmol/L and the intervention time for 72 h suggested the best lipolysis effect.After treatment of Myric and sh-RNA,size of lipid droplets and TG content decreased significantly compared with those in myricetin group or transfection group(P<0.05).The protein expression of PLIN1 downregulated while that of ATGL and HSL protein upregulated significantly in treatment group compared with those in myricetin group or transfection group(P<0.05).Conclusions Combinant treatment decreases the expression of PLIN1,increases the expression of ATGL and HSL and accelerate the lipolysis of 3T3-L1 adipocytes.