目的:观察蒲黄总黄酮(PTF)对C2C12骨骼肌细胞β-arrestin-2基因及蛋白表达的影响。方法:根据不同处理方法,将分化成熟的细胞分为对照组和PTF组。干预处理16h后,予或不予胰岛素(INS)刺激30min,实时定量PCR检测β-arrestin-2 mRNA表达,免疫印迹法检测β-arrestin-2蛋白表达,免疫共沉淀法检测β-arrestin-2信号复合物。结果:PTF组β-arrestin-2 mRNA表达水平是对照组的0.96倍,差异无统计学意义;但PTF组β-arrestin-2蛋白表达水平比对照组升高0.65倍,而PTF+INS组也比INS组升高0.66倍,有显著性差异(P〈0.01);与INS组相比较,PTF促进了β-arrestin-2信号复合物的形成。结论:PTF提高C2C12骨骼肌细胞β-arrestin-2蛋白表达和促进β-arrestin-2信号复合物形成,这可能是PTF改善胰岛素抵抗的机制之一。
Objective: To investigate the effects of Pollen Typhae total flavone (PTF) on the gene and protein expression of β -arrestin-2 in C2C12 myotubes. Methods: Differentiated cells were divided into the control group and the PTF group based on different treatment. Sixteen hours after treatment, the cells were incubated with or without insulin for 30 min. The β -arrestin-2 mRNA expression was assayed by Real-time PCR, protein level of β-arrestin-2 and its signal complex were detected by lmmunoblot and Co-immunoprecipitation respectively. Results: β -arrestin-2 mRNA level in the PTF group was 0.96 fold of the control group without statistical significance. But protein level of β -arrestin-2 in the PTF group was significantly increased by 0.65 fold when compared to the control group, while the PTF+insulin group by 0.66 fold (P〈0.01) when compared to the insulin group. Moreover, PTF promoted the formation of the β -arrestin-2 signal complex when compared to the insulin group. Conclusion: PTF enhanced the protein expression of β -arrestin-2 and the formation of the β -arrestin-2 signal complex, suggesting a mechanism of PTF in relieving insulin resistance.