生长激素、肝细胞生长因子和烟酰胺对人胎胰岛细胞体外增殖的影响

Effects of GH,HGF and NIC on proliferation and function of human fetal pancreatic islets in culture

目的 :探讨生长激素 (GH) ,肝细胞生长因子 (HGF)和烟酰胺 (NIC)对体外培养人胎胰岛细胞增殖的影响。方法 :胶原酶消化法制胰岛样细胞团 (ICCs) ,接种于 2 4孔培养板培养 ,在实验组中分别加入GH(1 0 0 μg/L) ,HGF(2 5 μg/L)和NIC(1 0mmol/L)及其组合 ,同时设空白组为对照 ,间日调换培养液并测定胰岛素分泌量 ,于培养 6d末测定胰岛细胞内胰岛素含量 ,收集细胞计数 ,观察细胞有丝分裂相及胰岛细胞分布情况。结果 :①在人胎胰岛细胞体外培养第 2d、第 4d、第 6d胰岛素分泌量各实验组均高于空白组 (P <0 .0 1 ) ,以GH +HGF +NIC组最高 ,GH +HGF +NIC组与GH +NIC组、GH +HGF组、HGF +NIC组相比 ,差异无统计学意义 ,但与GH组、HGF组、NIC组比较 ,差异有统计学意义 (P <0 .0 5 )。②各组胰岛细胞内胰岛素含量与胰岛样细胞团数均呈正相关 ,r均大于 0 .80 (P <0 .0 5 )。结论 :GH、HGF、NIC均能显著促进体外培养的人胎胰岛细胞的增殖 ,胰岛素含量及分泌量增加 ,3者有协同作用。

Aim:To study the effects of GH, HGF and NIC on the proliferation and function of human fetal pancreatic islets in culture.Methods:Islet-like cell clusters were isolated from human fetal pancreases by the collagenase and seeded into 24-well cell culture plastic slides (1 000 ICCs/well) in RPMI-1640 medium with one of the factors: GH,HGF and NIC or the mixture of both or all of them. The medium was changed every two days,the content of insulin in the medium was tested. After six days the cells were collected, dyed with DTZ, and the content of insulin in the cells was determined.Results:The number of the human fetal islet βcells and the content and the secretion of insulin were markedly promoted in all the experimental groups and even more significant in the experimental group containing GH+HGF+NIC by the end of the 2nd day, the 4th day and the 6th day.Conclusion: GH, HGF, and NIC could markedly promote the replication, insulin biosynthesis and secretion of human fetal islet β cells in vitro, and the mixture of all would gain the maximal effects.