论文部分内容阅读
目的:探讨亮氨酸拉链蛋白(GILZ)对3T3-L1前脂肪细胞增殖、分化的作用及对脂肪细胞分化相关标志基因PPARγ2、C/EBPa、LPL和FAS mRNA的表达。方法:采用MTT法检测GILZ稳定表达3T3-L1细胞从D1到D11细胞的增殖情况。油红O染色观察GILZ过表达对脂肪细胞分化和甘油三酯相对含量的影响。实时荧光定量PCR法检测脂肪细胞分化相关标志基因PPARγ2、C/EBPa、LPL和FAS的mRNA表达。结果:与转染Pc DNA3空载体的对照组相比,GILZ过表达组的细胞增殖差异无统计学意义(P>0.05)。经MID诱导后,与对照组相比,GILZ过表达组的桔红色细胞显著减少。脂肪细胞内甘油三酯相对含量也显著降低(0.365±0.012 vs 0.181±0.014,P<0.001)。分化过程中GILZ过表达的3T3-L1细胞内脂肪细胞分化基因PPARγ2、C/EBPa、LPL和FAS的mRNA表达显著降低(分化第9天时的相对表达分别为11.447±0.831 vs 1.173±0.290,17.700±0.915 vs 1.557±0.384,67.057±5.288 vs 9.467±3.406,40.946±3.968 vs 4.967±1.091,P<0.001)。结论:GILZ对前脂肪细胞的增殖没有明显的影响。GILZ过表达可显著性抑制PPARγ2,C/EBPa,LPL和FAS的mRNA表达,表明GILZ可能通过下调脂肪细胞分化转录因子PPARγ2,C/EBPa的表达而抑制脂肪细胞特异性基因LPL和FAS的表达,进而抑制脂肪细胞的分化。
Objective: To investigate the effect of leucine zipper protein (GILZ) on the proliferation and differentiation of 3T3-L1 preadipocytes and the expression of PPARγ2, C / EBPa, LPL and FAS mRNAs related to adipocyte differentiation. Methods: MTT assay was used to detect the proliferation of 3T3-L1 cells from D1 to D11 stably expressing GILZ. Oil red O staining observed GILZ overexpression on adipocyte differentiation and triglyceride relative content. Real-time fluorescence quantitative PCR was used to detect mRNA expression of PPARγ2, C / EBPa, LPL and FAS in adipocyte differentiation related genes. RESULTS: There was no significant difference in cell proliferation between GILZ overexpression group and control group transfected with pcDNA3 vector (P> 0.05). After induction by MID, the number of orange-red cells in GILZ overexpression group was significantly reduced compared with the control group. The relative content of triglycerides in adipocytes also decreased significantly (0.365 ± 0.012 vs 0.181 ± 0.014, P <0.001). The mRNA expression of adipocyte differentiation genes PPARγ2, C / EBPa, LPL and FAS in GILZ-overexpressed 3T3-L1 cells was significantly decreased during differentiation (relative expression at day 9 of differentiation was 11.447 ± 0.831 vs 1.173 ± 0.290 and 17.700 ± 0.915 vs 1.557 ± 0.384, 67.057 ± 5.288 vs 9.467 ± 3.406, 40.946 ± 3.968 vs 4.967 ± 1.091, P <0.001). Conclusion: GILZ has no obvious effect on the proliferation of preadipocytes. GILZ overexpression significantly inhibited the mRNA expression of PPARγ2, C / EBPa, LPL and FAS, indicating that GILZ may inhibit the expression of adipocyte-specific genes LPL and FAS by down-regulating the expression of adipocyte differentiation transcription factors PPARγ2 and C / EBPa, Which in turn inhibits adipocyte differentiation.