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目的探讨胰岛素对稳定表达人脂联素3T3-L1细胞PPARγ2mRNA表达的影响。方法重组脂联素真核表达质粒(pcDNA3.1+-hADPN)脂质体法稳定转染3T3-L1细胞。用胰岛素(500ng/mL)处理未转染、转染空载载体、转染pcDNA3.1+-hADPN的3T3-L1细胞,RT-PCR检测PPARγ2mRNA的表达量。结果①稳定转染了pcDNA3.1+-hADPN的未分化和已分化3T3-L1细胞,PPARγ2表达量较未转染组明显增加(P<0.01)。②胰岛素可明显抑制PPARγ2mRNA的表达(P<0.05)。③稳定转染pcDNA3.1+-hADPN可改善胰岛素抑制作用(P<0.05)。结论稳定转染pcDNA3.1+-hADPN可明显增加未分化和已分化的3T3-L1细胞PPARγ2mRNA表达。胰岛素抑制PPARγ2mRNA表达,转染pcDNA3.1+-hADPN可改善胰岛素抑制作用。
Objective To investigate the effect of insulin on PPARγ2 mRNA expression in human adiponectin-3T3-L1 cells stably. Methods Recombinant adiponectin eukaryotic expression plasmid (pcDNA3.1 + -hADPN) liposome method stably transfected 3T3-L1 cells. The untransfected and transfected 3T3-L1 cells transfected with empty vector and pcDNA3.1 + -hADPN were treated with insulin (500ng / mL). The expression of PPARγ2 mRNA was detected by RT-PCR. Results ① The expression of PPARγ2 in undifferentiated and differentiated 3T3-L1 cells stably transfected with pcDNA3.1 + -hADPN was significantly higher than that in untransfected cells (P <0.01). ② Insulin can significantly inhibit the expression of PPARγ2mRNA (P <0.05). ③ Stable transfected pcDNA3.1 + -hADPN can improve insulin inhibition (P <0.05). Conclusion Stably transfected pcDNA3.1 + -hADPN can significantly increase PPARγ2 mRNA expression in undifferentiated and differentiated 3T3-L1 cells. Insulin inhibits PPARγ2 mRNA expression. Transfection of pcDNA3.1 + -hADPN can improve insulin inhibition.