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目的探讨骨形态发生蛋白7(bone morphorgenetic protein 7,BMP7)对胰岛细胞胰岛素分泌的影响。方法Western blot检测小鼠胰岛细胞株MIN6中3种BMP7的Ⅰ型受体BMPR1A、BMPR1B、ACVR1A的表达情况。将MIN6细胞分为4组:PBS对照组,重组人BMP7(rhBMP7)低、中、高剂量组(rhBMP7终浓度分别为10、50、100 ng/ml),每组设5个复孔。ELISA检测各组细胞培养基中的胰岛素分泌情况。激光共聚焦显微镜检测细胞内钙离子浓度变化。结果BMPR1A、BMPR1B、ACVR1A均表达于MIN6细胞。PBS对照组及rhBMP7低、中、高剂量组的胰岛素浓度分别为(0.42±0.11)、(0.63±0.22)、(0.82±0.19)、(1.24±0.26)ng/(ml.μg)。与对照组相比,rhBMP7低、中、高剂量组的胰岛素分泌均明显增加(P<0.05)。PBS对照组及rhBMP7低、中、高剂量组的细胞内钙离子浓度增加分别为(3.5±2.1)%、(4.2±2.3)%、(5.1±3.5)%、(4.9±2.7)%,4组间差异不显著(P>0.05)。结论 BMP7可显著促进胰岛细胞的胰岛素分泌,其作用机制可能是调控非钙离子浓度依赖的胞吐环节。
Objective To investigate the effect of bone morphogenetic protein 7 (BMP7) on insulin secretion in pancreatic islet cells. Methods Western blot was used to detect the expression of BMPR1A, BMPR1B and ACVR1A in three kinds of BMP7 in mouse pancreatic islet cell line MIN6. MIN6 cells were divided into 4 groups: PBS control group, recombinant human BMP7 (rhBMP7) low, medium and high dose groups (rhBMP7 final concentrations of 10, 50, and 100 ng / ml). ELISA assay of insulin secretion in each group of cell culture medium. Laser confocal microscopy was used to detect intracellular calcium concentration changes. Results BMPR1A, BMPR1B and ACVR1A were all expressed in MIN6 cells. The insulin concentrations of PBS control group and rhBMP7 low, medium and high dose groups were (0.42 ± 0.11), (0.63 ± 0.22), (0.82 ± 0.19), (1.24 ± 0.26) ng / (ml.μg) respectively. Compared with the control group, the insulin secretion of rhBMP7 low, medium and high dose groups were significantly increased (P <0.05). (3.5 ± 2.1)%, (4.2 ± 2.3)%, (5.1 ± 3.5)%, (4.9 ± 2.7)%, 4 (4 ± 2)% respectively in PBS control group and rhBMP7 low, medium and high dose groups There was no significant difference between groups (P> 0.05). Conclusion BMP7 can significantly promote the insulin secretion of pancreatic islet cells. Its mechanism may be that it regulate the exocytosis of non-calcium ion concentration-dependent.