不同浓度脂联素对HepG2细胞内甘油三酯含量的影响及其机制

Influence of adiponectin with different doses on triglyceride contents in HepG2 cells and its mechanism

目的:观察不同浓度脂联素对HepG2细胞内载脂蛋白A5(apoA5)mRNA和蛋白表达水平及甘油三酯(TG)含量的影响,探讨脂联素与TG之间的剂量依赖关系,为研究脂联素降低TG含量的机制奠定基础。方法:将正常生长的HepG2细胞分为对照组、低剂量脂联素组、中剂量脂联素组和高剂量脂联素组,分别用不同浓度(0、12.5、25.0和50.0mg.L-1)脂联素作用24h,酶法检测各组HepG2细胞内TG含量,实时荧光定量PCR检测HepG2细胞内apoA5mRNA表达水平,Western blotting法检测HepG2细胞内apoA5mRNA蛋白表达水平。结果:与对照组比较,脂联素各组HepG2细胞内TG含量均明显降低(P<0.05);与低剂量脂联素组比较,中、高剂量脂联素组HepG2细胞内TG含量均明显降低(P<0.05),但高剂量组与中剂量组比较差异无统计学意义(P>0.05)。与对照组比较,脂联素各组HepG2细胞内apoA5mRNA和蛋白表达均明显增加(P<0.05);与低剂量脂联素组比较,中、高剂量脂联素组HepG2细胞内apoA5mRNA和蛋白表达均明显增加(P<0.05),但高剂量组与中剂量组比较差异无统计学意义(P>0.05)。结论:脂联素能降低HepG2细胞TG含量,并呈剂量依赖关系,提示脂联素降低肝脏中TG含量可能与其增加apoA5表达有关。

Objective To observe the influence of adiponectin with different doses on apolipoprotein A5（apoA5）mRNA and protein expression levels and triglyceride（TG） content in HepG2 cells and to investigate the relationship between adiponectin and TG and to establish foundation for study on the mechanism of adiponectin in decreasing TG content.Methods HepG2 cells were divided into control group,low dose adiponectin group,middle dose adiponectin group and high dose adiponectin group.The HepG2 cells in various groups were treated with different doses of adiponectin（0,12.5,25.0 and 50.0 mg·L-1）for 24 h,then the TG contents in HepG2 cells in various groups were detected with enzyme method.The expression levels of apoA5 mRNA in HepG2 cells were detected with real time fluorescence quantitative PCR method.The expression levels of apoA5 protein in HeG2 cells were detected with Western blotting method.Results Compared with control group,the TG contents in HepG2 cells in different doses of adiponectin groups were decreased obviously（P0.05）.Compared with low dose group,the TG contents in HepG2 cells in middle dose group and high dose group were decreased obviously（P0.05）,but there was no significant difference between high dose group and middle dose group（P0.05）.Compared with control group,the gene and protein expression levels of apoA5 in HepG2 cells in different doses of adiponectin groups were increased obviously（P0.05）.Compared with low dose group,the gene and protein expression levels of apoA5 in HepG2 cells in middle dose group and high dose group were increased obviously（P0.05）,but there was no significant difference between high dose group and middle dose group（P0.05）.Conclusion Adiponectin may decrease the TG content in HepG2 cells in an dose-dependent manner.It indicats that the mechanism of adiponectin in decreasing TG content may be related to increasing the expression of apoA5 in HepG2 cells.