中国组织工程研究

中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (24): 4481-4485.doi: 10.3969/j.issn.1673-8225.2012.24.023

• 组织构建细胞学实验 cytology experiments in tissue construction • 上一篇    下一篇

人脐静脉内皮细胞胰岛素抵抗细胞模型的建立

肖维民1，姚尚龙1，武庆平1，刁 波2，周 芳2，王 珏1，戴中亮1，伍 静1   

  1. 1华中科技大学同济医学院附属协和医院麻醉科，湖北省武汉市430030；
    2解放军广州军区武汉总医院动物实验中心，广东省广州市 430070
  • 收稿日期:2012-03-02 修回日期:2012-05-02 出版日期:2012-06-10 发布日期:2013-11-05
  • 通讯作者: 肖维民，男，1971年生，湖北省钟祥市人，汉族，1994年华中科技大学同济医学院毕业，博士，副主任医师，主要从事全麻机理及脏器功能保护研究。 xiaoweimin199968@163.com 伍静，博士，主治医师，华中科技大学同济医学院附属协和医院麻醉科，湖北省武汉市 430030 icecream555@163.com
  • 作者简介:肖维民，男，1971年生，湖北省钟祥市人，汉族，1994年华中科技大学同济医学院毕业，博士，副主任医师，主要从事全麻机理及脏器功能保护研究。 xiaoweimin199968@163.com
  • 基金资助:

    湖北省自然科学基金资助项目(2006AA301B55)。课题名称：围手术期血管内皮胰岛素抵抗的发生及机制研究

Establishment of insulin resistance models of human umbilical vein endothelial cells

Xiao Wei-min1, Yao Shang-long1, Wu Qing-ping1, Diao Bo2, Zhou Fang2, Wang Jue1, Dai Zhong-liang1, Wu Jing1   

  1. 1Department of Anesthesiology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, Hubei Province, China;
    2Department of Medical Experiment, Wuhan General Hospital of Guangzhou Military Region, Wuhan 430070, Hubei Province, China
  • Received:2012-03-02 Revised:2012-05-02 Online:2012-06-10 Published:2013-11-05
  • Contact: Xiao Wei-min, Department of Anesthesiology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, Hubei Province, China xiaoweimin199968@163.com Wu Jing, Doctor, Attending physician, Department of Anesthesiology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, Hubei Province, China icecream555@163.com
  • About author:Xiao Wei-min☆, Doctor, Associate chief physician, Department of Anesthesiology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, Hubei Province, China xiaoweimin199968@163.com

PDF

432

被引次数

3

摘要：

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(50，40，30，20，10，1，0.1，0.01 U/L)的DMEM培养基与人脐静脉内皮细胞共同培养12，24，36，48，72 h，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40 U/L，其作用时间大于48 h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20 U/L，作用时间小于36 h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30 U/L的胰岛素刺激48 h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30 U/L胰岛素的培养基培养48 h的人脐静脉内皮细胞细胞可产生胰岛素抵抗。

关键词: 脐静脉内皮细胞, 胰岛素抵抗, 细胞活性, 细胞模型, 组织构建

Abstract:

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.

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