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目的先前研究已显示resistin结合多肽(RBP)能拮抗resistin对脂肪细胞脂质代谢和内分泌功能的调节作用。本研究试图阐明RBP对胰岛β细胞株RINm5F分泌功能的影响。方法以不同水平RBP(10-10,10-11,10-12mol/L)与RINm5F共培养30min、60min、2h后取上清,ELISA法测其胰岛素,RT-PCR法检测细胞中葡萄糖转运子2(GLUT2)mRNA表达水平,Western blotting法检测细胞中GLUT2的蛋白水平,并采用FURA-3/AM钙荧光染色法检测胰岛素分泌的重要启动因素:细胞内钙水平。结果RBP在10-12mol/L时干预2h,不影响胰岛β细胞株RINm5F的细胞活性,但能显著刺激胰岛素分泌。RBP为10-12mol/L时干预2h,细胞内钙明显增多。在Resistin刺激下,GLUT2mRNA水平和蛋白水平均显著上调。结论RBP能促进RINm5F细胞株胰岛素的分泌,其可能机制是RBP上调了GLUT2的表达,进而增加了细胞内钙的水平,最终导致胰岛素分泌增加。
PURPOSE Previous studies have shown that resistin binding polypeptide (RBP) antagonizes the modulatory effect of resistin on adipocyte lipid metabolism and endocrine function. This study sought to elucidate the effect of RBP on the secretion of pancreatic β-cell RINm5F. Methods Different levels of RBP (10-10,10-11,10-12 mol / L) and RINm5F were co-cultured for 30min, 60min, 2h after the supernatant, ELISA measured insulin, RT-PCR assay glucose transporter 2 (GLUT2) mRNA was detected by Western blotting. The protein level of GLUT2 in cells was detected by Western blotting. The level of intracellular calcium was measured by FURA-3 / AM calcium fluorescence staining. Results RBP intervention at 10-12 mol / L for 2 h did not affect the cellular activity of pancreatic β-cell line RINm5F, but significantly stimulated insulin secretion. RBP 10-12mol / L intervention 2h, intracellular calcium increased significantly. Under Resistin stimulation, both GLUT2 mRNA and protein levels were significantly up-regulated. Conclusion RBP can promote the insulin secretion in RINm5F cell line. The possible mechanism is that RBP up-regulates the expression of GLUT2, which in turn increases the level of intracellular calcium and finally leads to the increase of insulin secretion.