人脐静脉内皮细胞胰岛素抵抗细胞模型的建立

背景:有研究表明血管内皮可能是胰岛素抵抗发生的首要环节。目的:采用高浓度胰岛素体外诱导培养法建立人脐静脉内皮细胞胰岛素抵抗模型。方法:以人脐静脉内皮细胞系为研究对象,应用含不同浓度胰岛素(50,40,30,20,10,1,0.1,0.01U/L)的DMEM培养基与人脐静脉内皮细胞共同培养12,24,36,48,72h,并设置正常组,光学显微镜下观察细胞形态学变化;MTT法测定各组细胞活性;用葡萄糖氧化酶-过氧化物酶法法鉴定胰岛素抵抗细胞模型。结果与结论:与正常组比较,当胰岛素浓度大于40U/L,其作用时间大于48h时,细胞形态受损严重,细胞活性显著下降(P<0.01);当胰岛素浓度小于20U/L,作用时间小于36h时细胞形态无明显损伤(P>0.05)。葡萄糖氧化酶-过氧化物酶法实验表明,在30U/L的胰岛素刺激48h条件下,培养液中残存葡萄糖浓度显著高于正常组细胞(P<0.01)。证实,用含30U/L胰岛素的培养基培养48h的人脐静脉内皮细胞细胞可产生胰岛素抵抗。

Establishment of insulin resistance models of human umbilical vein endothelial cells

BACKGROUND:Studies have shown that vascular endothelium may be the first link of insulin resistance.OBJECTIVE:To establish insulin resistance models of human umbilical vein endothelial cells(HUVECs) induced by hyperinsulinsm.METHODS:HUVECs were chosen as the research object and were incubated in Dulbecco’s modified Eagle’s medium with different concentrations of insulin(50,40,30,20,10,1,0.1,0.01 U/L) for 12,24,36,48,72 hours.Normal group was set.The change of cellular form was observed under optical microscope,and the cell viability in each group was determined by MTT.Glucose oxidizes peroxides was done to evaluate whether the insulin resistance model was successfully prepared.RESULTS AND CONCLUSION:Compared with the normal group,the morphology of HUVECs incubated in Dulbecco’s modified Eagle’s medium containing insulin more than 40 U/L for more than 48 hours was badly damaged and the cell viability decreased significantly(P < 0.01).When the concentration of insulin was less than 20 U/L,and incubated time was less than 36 hours,no significantly damage was found in the cellular form,the cell viability decreased but not significantly(P > 0.05).The results of glucose oxidizes peroxides showed that the glucose content in the culture medium in which HUVECs were incubated with 30 U/L insulin for 48 hours was higher than that in the normal group significantly(P﹤0.01).These findings indicate that HUVECs incubated in the culture medium containing 30 U/L insulin for 48 hours can present with insulin resistance.