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目的 观察游离脂肪酸对大鼠骨骼肌细胞葡萄糖转运蛋白 4(GLUT4)和胰岛素信号传导蛋白Grb2及ERK2的影响。方法 分离、培养新生Sprague Dawley大鼠骨骼肌细胞 ,分别与软脂酸(0 2 5mmol/L)或油酸 (0 12 5mmol/L)孵育 12、2 4、36h ,提取蛋白后用Western印迹法检测GLUT4、Grb2和ERK2的蛋白水平 ;用斑点印迹杂交法检测骨骼肌细胞内GLUT4RNA含量的变化。结果 经软脂酸和油酸孵育 12、2 4、36h后 ,大鼠骨骼肌细胞GLUT4的蛋白和RNA水平均显著降低 (P <0 0 5 ) ;同时 ,Grb2和ERK2的蛋白水平也明显下降 (P <0 0 5 )。结论 游离脂肪酸可抑制GLUT4的基因及蛋白表达和降低胰岛素信号传导蛋白Grb2和ERK2的蛋白表达水平 ,这可能会影响胰岛素的信号传导作用和骨骼肌的葡萄糖摄取 ,引起骨骼肌的胰岛素抵抗
Objective To observe the effects of free fatty acids on glucose transporter 4 (GLUT4) and insulin signaling proteins Grb2 and ERK2 in rat skeletal muscle cells. Methods Skeletal muscle cells of neonatal Sprague Dawley rats were isolated and incubated with palmitic acid (0 2 5 mmol / L) or oleic acid (0 12 5 mmol / L) for 12, 24 and 36 h, respectively. The protein levels of GLUT4, Grb2 and ERK2 were detected; the changes of GLUT4RNA content in skeletal muscle cells were detected by dot blot hybridization. Results After 12, 24 and 36 h of incubation with palmitic acid and oleic acid, the protein and RNA levels of GLUT4 in rat skeletal muscle cells were significantly decreased (P <0.05). Meanwhile, the protein levels of Grb2 and ERK2 were significantly decreased (P <0 05). Conclusions Free fatty acids can inhibit the gene and protein expression of GLUT4 and decrease the protein expression of insulin signaling proteins Grb2 and ERK2, which may affect insulin signaling and glucose uptake in skeletal muscle and cause insulin resistance in skeletal muscle