Visfatin基因表达下调对小鼠脂肪细胞及肝细胞脂代谢的影响

Effects of visfatin down-regulation on lipid metabolism in 3T3-L1 cells and hepa1-6 cells

目的探讨Visfatin基因表达抑制对小鼠脂肪细胞及肝细胞脂代谢相关基因的影响。方法构建并筛选具有最佳抑制率的pGene-sil-Visfatin表达载体,转染小鼠3T3-L1脂肪细胞及Hepa1-6肝细胞,Western印迹法测定两种细胞的Visfatin蛋白水平,实时荧光定量PCR法测定Vis-fatin、脂代谢相关基因的mRNA表达水平,油红O染色法测定脂肪细胞内甘油三酯(TG)含量变化。结果 pGenesil-visfatin可有效抑制两种细胞Vis-fatin蛋白表达。脂肪细胞转染pGenesil-Visfatin后,脂代谢相关基因PPARα/γ,HSL,ATGL,AP2,SREBP1,FAS,ACC mRNA表达均下降(其中PPARγ与ACC P<0.05,余P<0.001),TG含量降低,但无统计学差异;肝细胞转染后,PPARα,SREBP1,FAS以及SREBP2,HMGCR,LDLr mRNA表达显著降低(P<0.001),而HSL,ATGL与ACC变化无统计学意义(P>0.05)。结论 Visfatin基因表达下调可能对细胞脂质代谢具有调节作用。

Objective To investigate the effects of visfatin-shRNA on lipid metabolism in 3T3-L1 ceils and Hepal-6 cells. Meth-ods pGenesil-visfatin was transfected into differentiated 3T3-L1 cells and Hepal-6 cell respectively. The visfatin protein was measured by western blot. The mRNA levels of genes involved in lipid metabolism were measured in adipocytes and hepatic cells by real time PCR. TG content in adipocytes was detected by colorimetric assay at 490 nm. Results The protein content of visfatin was inhibited effectively by pGenesil-visfatin recombinant plasmid. In pGenesil-visfatin transfected 3T3-L1 cells, down-regulation of visfatin reduced the lipid metabolism related gene expression of PPARα/γ, HSL, ATGL, AP2, SREBP1, FAS, ACC （P 〈0. 05, P 〈0. 001 ）. Intraeellular TG content was de-creased, but there was no significant difference. In Hepal-6 cells, down-regulation of visfatin by pGenesil-visfatin transfeetion reduced mR-NA level of PPARα, SREBP1, FAS, SREBP2, HMGCR, LDLr significantly （ all P 〈 0. 001 ） , while the expression of HSL, ATGL and ACC hadn＇t changed significantly （P 〉 0. 05）. Conclusions Down-regulation of visfatin can regulate the gene expression involved in lipid metabolism.