原代培养骨骼肌细胞胰岛素抵抗模型的建立

Establishment of Insulin Resistance Model in Primary Culture Skeletal Muscle Cell

目的:观察高脂负荷在胰岛素抵抗形成中的意义。方法:用不同浓度棕榈酸、胰岛素分别培养骨骼肌细胞2h、6h、12h、24h,对棕榈酸诱导组的一部分细胞给予1×10^(-7)M胰岛素刺激2h,用血糖检测试剂盒(GOD-POD)检测各组培养液中的葡萄糖含量,研究不同浓度棕榈酸、胰岛素对骨骼肌细胞摄取葡萄糖的影响,观察胰岛素的生理功效的变化。结果:0.6mM棕榈酸诱导12h以上或者5×10^(-7)M胰岛素诱导24h后,培养液中的葡萄糖浓度比正常组高且有显著性差异,表明细胞的糖代谢能力降低。经1×10^(-7)M胰岛素刺激2h后的棕榈酸诱导组,培养液中葡萄糖的浓度与未经胰岛素刺激的棕榈酸诱导组相比无显著差异,胰岛素的生理功效降低,证实棕榈酸诱导组细胞已对胰岛素产生耐受。结论:高脂或高胰岛素条件均可诱导原代骨骼肌细胞胰岛素抵抗模型。

Objective： To observe the significance of high palmitic acid （PA） in insulin resistance formation. Methods： Skeletal muscle cell was treated with different concentration of PA or insulin for 2 h, 6 h, 12 h, and 24 h. One part of cells, which had been treated by PA, were additional incubated with l×10^-7 M insulin for 2 h. The concentration of glucose was determined by Glucose Detection Kit （GOD-POD）. Then, the effect of different concentration of PA or insulin on glucose uptake was investigated in skeletal muscle cell. Moreover, the efficacy of insulin was also estimated. Results： The concentration of glucose in the induced group was significantly higher than that in the control group when the cells were incubated with palmitic acid （PA, ≥ 0.6 raM） for 12 h, or with 5×10^-7 M insulin for 24 h., and which indicated that the metabolic capability of glucose was depressed. The concentration of glucose in the additional induced group with insulin had no difference with that in no additional induced group, which also indicated that the efficacy of insulin was cut down. Conclusion： Insulin resistance model in primary skeletal muscle cell can be established by culturing the primary skeletal muscle cell in high concentration of PA or insulin.