骨形态发生蛋白7对胰岛素分泌的影响及机制研究.docVIP

骨形态发生蛋白7对胰岛素分泌的影响及机制研究 姜友昭，曾 俊，陈 兵（400038重庆，第三军医大学西南医院内分泌科） [摘要]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。然后将MIN6细胞分为4组：①PBS对照组；②重组人BMP7（rhBMP7）低剂量组（rhBMP7终浓度为10 ng/ml ）；③rhBMP7中剂量组（rhBMP7终浓度为50 ng/ml）；④rhBMP7大剂量组（rhBMP7终浓度为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、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（P0.05）。而PBS对照组、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（P0.05）。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。 [关键词]骨形态发生蛋白7；胰岛素分泌；MIN6细胞 The effect of bone morphorgenetic protein 7 on insulin secretion and its mechanism Jiang You-zhao， Zeng Jun，Chen Bing（Department of Endocrinology， Southwest Hospital，Third Military Medical University， Chongqing， 400038， China ） [Abstract] Objective To investigate the effect of bone morphorgenetic protein 7（BMP7）on insulin secretion in pancreatic beta cells and its mechanism. Methods The expression of three type I receptors of BMP7 was detected by western blot in mouse pancreatic beta cell line MIN6. Then the MIN6 cells were divided into four groups： ①PBS control group；②group dealt with low-dose rhBMP7（final concentrtion of rhBMP7 was 10 ng/ml）；③group dealt with medium-dose rhBMP7（final concentrtion of rhBMP7 was 50 ng/ml）；④group dealt with high-dose rhBMP7（final concentrtion of rhBMP7 was 100 ng/ml）. Of each gpoups， five independent experiments run in duplicate. Then the insulin level of the media was deteceted by ELISA and the intracellular concentration of calcium was determined by laser confocal microscope. Results BMPR1A、BMPR1B、ACVR1A were all expressed in the MIN6 cells. The insulin concentration of PBS control group、low-dose， medium-dose and high-dose rhBMP7 group was 0.42±0.11、0.63±0.22、0.82±0.0.19、1.24±0.26 ng/（ml·μg）， respectively. The insulin concentratio