2,2',4,4'-四溴联苯醚对小鼠3T3-L1细胞分化的影响研究

Effects of 2,2’,4,4’-tetrabromodiphenyl ether on differentiation of mouse 3T3-L1 cells

目的研究2,2’,4,4’-四溴联苯醚(BDE-47)对小鼠胚胎成纤维细胞3T3-L1分化的影响,为揭示环境肥胖因素的作用规律提供依据。方法将3T3-L1细胞分为5组BDE-47干预组(25、18.75、12.5、7.5、2.5μmol/L)、阳性对照组(加1μmol/L 2,4-噻唑烷二酮)和阴性对照组(加0.1%二甲基亚砜),诱导分化第8 d油红O染色处理并检测吸光度值(OD)观察脂肪细胞脂滴聚集情况;采用三酰甘油(TG)酶法测定细胞培养液的TG含量;采用RT-PCR法测定脂联素和过氧化物酶体增殖子激活受体γ(PPARγ)的mRNA表达。结果7组3T3-L1细胞油红O染色阳性面积、OD值、TG含量及脂联素和PPARγ的mRNA表达量差异均有统计学意义(P<0.05)。不同浓度BDE-47组的油红O染色阳性面积、OD值均高于阴性对照组(P<0.05)。18.75μmol/L BDE-47组TG含量高于阴性对照组(P<0.05)。25、18.75、12.5、7.5μmol/L BDE-47组和阳性对照组PPARγ的mRNA相对表达量均高于阴性对照组(P<0.05);12.5μmol/L BDE-47组PPARγ的mRNA相对表达量均高于25、18.75、7.5、2.5μmol/L BDE-47组和阳性对照组(P<0.05)。12.5、7.5μmol/L BDE-47组和阳性对照组脂联素的mRNA相对表达量均高于阴性对照组(P<0.05);12.5μmol/L BDE-47组脂联素的mRNA相对表达量均高于25、18.75、2.5μmol/L BDE-47组(P<0.05)。不同浓度BDE-47组的PPARγ和脂联素mRNA表达量近似倒"U"型分布。结论BDE-47对小鼠3T3-L1细胞分化起促进作用,低浓度的BDE-47可能通过激活PPARγ活性诱导脂肪细胞分化。

Objective To investigate the effects of 2,2’,4,4’-tetrabromodiphenyl ether(BDE-47)on the differentiation of mouse embryonic fibroblasts-3T3-L1,so as to provide the basis for revealing the mechanism of environmental obesity factors.Methods The 3T3-L1 cells were divided into five BDE-47 intervention groups(25,18.75,12.5,7.5 and 2.5μmol/L),a positive control group(1μmol/L 2,4-thiazolidinedione)and a negative control group(0.1%dimethyl sulfoxide)for the induction of differentiation.The lipid droplet accumulation in adipocytes was observed by oil red O staining treatment and detection of optical desity(OD)on the eighth day of differentiation.Triglyceride(TG)content was measured using the histiocyte TG enzymatic assay kit.The mRNA expression of adiponectin and peroxisome proliferator activated receptor-γ(PPARγ)was measured by RT-PCR.Results The positive areas of oil red O staining,OD values,TG content and mRNA expression of adiponectin and PPARγin 3T3-L1 cells were significantly different among seven groups(P<0.05).The positive areas of oil red O staining and OD values in the BDE-47 groups with different concentrations were higher than those in the negative control group(P<0.05).The 18.75μmol/L BDE-47 group had higher TG levels than the negative control group(P<0.05).The mRNA expression of PPARγin the 25,18.75,12.5,and 7.5μmol/L BDE-47 groups and the positive control group was higher than that in the negative control group(P<0.05).The mRNA expression of PPARγin the 12.5μmol/L BDE-47 group was higher than that in the 25,18.75,7.5,2.5μmol/L BDE-47 group and the positive control group(P<0.05).The mRNA expression of adiponectin in the 12.5,7.5μmol/L BDE-47 group and the positive control group was higher than that in the negative control group(P<0.05).The mRNA expression of adiponectin in the 12.5μmol/L BDE-47 group was higher than that in the 25,18.75,2.5μmol/L BDE-47 group(P<0.05).The mRNA expression of PPARγand adiponectin in the different concentration groups of BDE-47 distributed like inverted"U"shape.Conclusion BDE-47 can promote the differentiation of 3T3-L1 preadipocytes.Low concentration of BDE-47 may induce adipocyte differentiation by activating PPARγ.