间断性高浓度胰岛素对HUECV304细胞iNOS蛋白表达的影响  

Influence of discontinuous high insulin on protein expression of iNOS in HUECV304

作　　者：黄凌云[1,2] 秦爱平 杨敏 刘江华[2] 全宏梅 廖斌 

机构地区：[1]湖南省老年医院,长沙410016 [2]南华大学附属第一医院,衡阳421001 [3]国家康复辅具研究中心附属康复医院,北京100049

出　　处：《右江医学》2015年第1期11-15,共5页Chinese Youjiang Medical Journal

基　　金：湖南省科技厅资助项目(编号:2007FJ4135)

摘　　要：目的研究体外培养的人脐静脉内皮细胞(HUECV304)在间断性高浓度胰岛素与持续性高浓度胰岛素作用下诱导型一氧化氮合酶(i NOS)蛋白的不同表达,从而进一步研究间断性高浓度胰岛素血症导致动脉粥样硬化的原因及发病机制。方法培养人脐静脉内皮细胞(HUECV304)为实验对象,研究间断性高浓度胰岛素对血管内皮细胞i NOS蛋白表达的影响。分为四组进行实验,即间断性高浓度胰岛素波动组(含100 nmol/L胰岛素的细胞培养液及含1 nmol/L胰岛素的细胞培养液轮换,每隔8小时更换一次)、持续性高浓度胰岛素组(含100 nmol/L胰岛素的细胞培养液)、含正常浓度胰岛素组(含1 nmol/L胰岛素的细胞培养液)、阴性对照组(不含胰岛素的细胞培养液);再依次更换新鲜条件细胞培养液(每次培育8小时后),四组细胞均培育72小时。采用免疫细胞化学法检测内皮细胞i NOS蛋白表达。结果四组内皮细胞经含不同浓度胰岛素的培养液培养72小时后,结果表明:正常浓度胰岛素组、间断性高浓度胰岛素组、持续性高浓度胰岛素组的内皮细胞i NOS蛋白的表达水平均不同程度地增高,分别为对照组的1.92倍(P<0.001)、4.52倍(P<0.001)、3.31倍(P<0.001),其中以间断性高浓度胰岛素组增高幅度尤为明显,高于持续性高浓度胰岛素组(P<0.001)。结论相对于持续性高浓度胰岛素,间断性高浓度胰岛素更能增强人脐静脉内皮细胞i NOS蛋白的表达,提示间断性高浓度胰岛素可能通过影响血管内皮细胞i NOS蛋白表达途径而损伤血管内皮的功能,从而促进糖尿病血管并发症的进一步发展。Objective To study influence of discontinuous and persistent high insulin on different protein expressions of induced nitric oxide synthase（ i NOS） in cultured human umbilical vascular endothelial cell（ HUECV304）,so as to explore causes and mechanism of atherosclerosis resulted from intermittent hyperinsulinemia. Methods HUECV304 was selected as the experimental subject,and influence of discontinuous and persistent high insulin on different protein expressions of i NOS were investigated. HUECV304 were divided into the following four groups： discontinuous high insulin fluctuate group（ with100 nmol / L and 1 nmol / L of insulin cell culture medium rotating every 8 hours）,persistent high insulin group（ with 100 nmol / L of insulin cell culture medium）,normal insulin group（ with 1 nmol / L of insulin cell culture medium） and negative control group（ without insulin cell culture medium）. Cell culture medium with fresh conditions were replaced in turn after culturing for every 8 hours. And endothelial cells of the four groups were cultured for 72 hours,and the protein expressions of i NOS were detected by immunocytochemistry. Results After the endothelial cells in the four groups were cultured for 72 hours by different concentrations of insulin cell culture medium,the results showed that the protein expressions levels of i NOS of the normal insulin group,the discontinuous high insulin fluctuate group and the persistent high insulin group increased in varying degrees,and they were 1. 92 times（ P 〈0. 001）,4. 52 times（ P 〈0. 001） and 3. 31 times（ P 〈0. 001） higher than that of the control group respectively. Among all of the groups,protein expression level of i NOS in the discontinuous high insulin fluctuate group improved more significantly,higher than that of the persistent high insulin group（ P 〈0. 001）. Conclusion Discontinuous high insulin can enhance protein expression level of i NOS in HUECVs more significantly compared with persistent high insulin,which prompts that

关 键 词：间断性高浓度胰岛素 人脐静脉内皮细胞 诱导型一氧化氮合酶 蛋白表达 

分 类 号：R587.1[医药卫生—内分泌]