不同浓度胰岛素对脐静脉内皮细胞形态和活性的影响  

Effects of Different Concentrations of Insulin on the Morphometry and Cytoactive of Human Umbilical Vein Endothelial Cells

作　　者：肖维民[1] 姚尚龙[1] 刁波[2] 周芳[2] 武庆平[1] 王珏[1] 戴中亮[1] 伍静[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院麻醉科,湖北武汉430061 [2]广州军区武汉总医院动物实验中心

出　　处：《华南国防医学杂志》2011年第4期280-283,共4页Military Medical Journal of South China

基　　金：湖北省自然科学基金重大项目(2006AA301B55)

摘　　要：目的观察不同浓度胰岛素对人脐静脉内皮细胞(human umbilical vein endothelial cell,HUVEC)形态和细胞活性的影响,用以筛选内皮细胞胰岛素抵抗模型。方法应用含不同浓度胰岛素(50,40,30,20,10,1,0.1,0.01U/L)的DMEM培养基与HUVEC共同培养6、12、24、36、48、72h;观察细胞形态学变化;四甲基偶氮唑盐比色法(tet-razolium colormetric assay,MTT)测定细胞活性。结果①细胞形态学变化:当胰岛素浓度小于10U/L,HUVEC生长加速;当胰岛素浓度为30U/L时,HUVEC出现核固缩;当胰岛素浓度达到50U/L时,HUVEC出现核碎裂和核溶解。②MTT法测定细胞活性:与0.01U/L组比较,当胰岛素浓度为50U/L时,细胞活性显著下降(P<0.01),当胰岛素浓度小于10U/L时,模型组细胞活性无统计学变化(P>0.05)。当作用时间超过36h,残留细胞数明显下降,细胞活性显著降低(P<0.01)。③析因设计资料的方差分析结果:不同胰岛素剂量组间差异有统计学意义(P<0.01);不同用药时间(胰岛素作用时间)组间差异有统计学意义(P<0.01);胰岛素剂量和用药时间之间交互效应有统计学意义(P<0.01);模型拟合效果理想,R2=0.928。交互效应轮廓图也提示胰岛素给药剂量和作用时间存在交互效应。结论胰岛素对人脐静脉内皮细胞的生长发育具有双向调控作用,胰岛素浓度30U/L、作用时间48h的有可能成为血管内皮细胞胰岛素抵抗的细胞模型。Objective By observing the effect of different concentrations of insulin on the morphometry and cytoactive of human umbilical vein endothelial cell（HUVEC） to select the insulin resistance model of HUVEC.MethodsDMEM with insulin of different concentrations（50,40,30,20,10,1,0.1,0.01 U/L） and HUVECs were cultivated together for 6,12,24,36,48 and 72 hours.The changes of cell morphology were observed and the cytoactive was tested by tetrazolium colormetric assay（MTT）.Results ①The changes of cell morphology.The growth of HUVEC sped up when the insulin concentration was less than 10 U/L.The karyopyknosis of HUVEC was observed at the concentration of 30 U/L and karyorrhexis and caryolysis were observeded at 50 U/L.②The cytoactive determined by MTT.Compared with that of the group with 0.01 U/L insulin,the cytoactive was decreased significantly in the group with 50 U/L insulin（P〈0.01）,and there was no significant difference in the group with less than 30 U/L insulin（P〈0.05）.When the action time was more than 36 hours,the number and cytoactive of residual cells were descended significantly（P〈0.01）.③ The ANOVA results of the factorial design.There were significant differences among the groups of different insulin concentrations（P〈0.01）,and of different medication times（the action time of insulin,P〈0.01）.The interactions between the dosage and medication were significant（P〈0.01） which was also ascertained by the contour map of interaction.A good fitness of the model was obtained（R^2=0.928）.Conclusion Insulin has bilateral regulation on the growth and development of HUVECs.The model of 30U/L insulin and 48 hours action time may be ideal for insulin resistance of vein endothelial cells.

关 键 词：胰岛素 脐静脉内皮细胞 胰岛素抵抗 

分 类 号：R587.1[医药卫生—内分泌]