Nesfatin-1对3T3-L1脂肪细胞糖代谢和自噬的影响

Effects of Nesfatin-1 on Glycometabolism and Autophagy in 3T3-L1 Adipocytes

为探讨厌食肽Nesfatin-1对脂肪细胞糖代谢和自噬的影响,本试验通过体外诱导分化3T3-L1前脂肪细胞,并在高糖状态下,以Nesfatin-1干预1 h之后,采用非放射性荧光法、ELISA、实时荧光定量PCR和Western blot分别检测脂肪细胞的葡萄糖摄取水平、丙酮酸含量、己糖激酶和磷酸果糖激酶活性以及自噬因子LC3、p62和Beclin-1 mRNA和蛋白表达量。结果显示,3T3-L1前脂肪细胞经过8 d诱导可达到完全分化状态;与对照组相比,经过Nesfatin-1处理后,3T3-L1脂肪细胞的葡萄糖摄取水平极显著降低(P<0.01),己糖激酶和磷酸果糖激酶的酶活性均非常显著地降低(P<0.001),p62 mRNA和蛋白表达量极显著下降(P<0.01),但丙酮酸含量、Beclin-1 mRNA以及LC3 mRNA和蛋白表达量差异不显著(P>0.05)。结果提示,Nesfatin-1不仅能有效降低3T3-L1脂肪细胞的糖消耗能力,还能上调脂肪细胞的自噬水平。

In order to explore the effects of anorexigenic peptide Nesfatin-1 on glycometabolism and autophagy in adipocytes,this study induced differentiation of 3T3-L1 preadipocytes in vitro.Under high glucose conditions,after 1 hour of Nesfatin-1 intervention,non-radioactive fluorescence assays,ELISA,Real-time fluorescence quantitative PCR,and Western blot were used to measure the glucose uptake level,pyruvate content,activities of hexokinase and phosphofructokinase,and the mRNA and protein expression levels of autophagy-related factors LC3,p62,and Beclin-1 in adipocytes.The results showed that 3T3-L1 preadipocytes reached complete differentiation after 8 days of induction.Compared with the control group,after Nesfatin-1 treatment,the glucose uptake level of 3T3-L1 adipocytes was very significantly reduced(P<0.01),the enzymatic activities of hexokinase and phosphofructokinase were extremely significant reduced(P<0.001),the mRNA and protein expression levels of p62 were very significantly decreased(P<0.01),while pyruvate content,Beclin-1 mRNA,LC3 mRNA and protein expression levels showed no significant differences(P>0.05).These results suggest that Nesfatin-1 not only effectively reduces glucose consumption in 3T3-L1 adipocytes but also upregulates the level of autophagy in these cells.