›› 2011, Vol. 31 ›› Issue (5): 543-.doi: 10.3969/j.issn.1674-8115.2011.05.004

• 论著(基础研究) • 上一篇    下一篇

晚期糖基化终末产物对巨噬细胞Nampt表达的影响

简蔚霞, 林 宁, 董 艳, 苏 青   

  1. 上海交通大学 医学院附属新华医院内分泌科, 上海 200092
  • 出版日期:2011-05-28 发布日期:2011-05-27
  • 通讯作者: 苏 青, 电子信箱: dr_suqing@yahoo.com.cn。
  • 作者简介:简蔚霞(1977—), 主治医师, 博士;电子信箱: jianweixia@yahoo.com。
  • 基金资助:

    高等学校博士学科点专项科研基金 (30900699);上海高校选拔培养优秀青年教师科研专项基金(jdy08084);上海交通大学医学院基金(2008XJ020)

Role of advanced glycation end products on expression of Nampt in macrophages

JIAN Wei-xia, LIN Ning, DONG Yan, SU Qing   

  1. Department of Endocrinology, Xinhua Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200092, China
  • Online:2011-05-28 Published:2011-05-27
  • Supported by:

    Scientific Research Foundation for Doctorate Program of College, 30900699;Foundation for Cultivation of Excellent Young Teachers in Shanghai Colleges, jdy08084;Shanghai Jiaotong University School of Medicine Foundation, 2008XJ020

摘要:

目的 探讨晚期糖基化终末产物(AGEs)对巨噬细胞尼克酰胺磷酸核糖转移酶(Nampt)表达的影响。方法 以人单核细胞株THP-1为体外细胞干预模型,经佛波酯诱导分化为巨噬细胞后,以0(阴性对照组)、50、100、250、500、1 000 μg/mL 的AGEs干预12 h,采用Western blotting方法检测细胞Nampt蛋白表达。以Western blotting检测结果选择合适浓度的AGEs处理巨噬细胞,分别于干预后0(阴性对照组)、3、6、12、24 h时点收集细胞,采用实时定量聚合酶链式反应(Real-Time PCR)技术和Western blotting方法检测巨噬细胞Nampt mRNA和蛋白表达。结果 在50~500 μg/mL的浓度范围内,AGEs处理组巨噬细胞Nampt 蛋白表达量均显著高于阴性对照组(P<0.01),且随干预浓度的升高呈现逐渐上升趋势,250 μg/mL和500 μg/mL AGEs处理组的干预效应最显著,设定后续实验中以250 μg/mL AGEs对巨噬细胞进行干预。250 μg/mL AGEs处理组巨噬细胞干预后各时点的Nampt mRNA和蛋白表达量均显著高于阴性对照组(P<0.01),且随干预时间的延长呈现逐渐上升趋势。结论 AGEs干预能促进巨噬细胞Nampt表达上调;提示AGEs可能部分通过Nampt途径激活巨噬细胞而参与糖尿病动脉粥样硬化的发生过程。

关键词: 晚期糖基化终末产物, 巨噬细胞, 尼克酰胺磷酸核糖转移酶

Abstract:

Objective To investigate the role of advanced glycation end products (AGEs) on expression of nicotinamide phosphoribosyltransferase (Nampt) in macrophages. Methods The macrophages were differentiated from human monocyte THP-1 by phorbol ester, and were treated with AGEs of 0 μg/mL (negative control group), 50 μg/mL, 100 μg/mL, 250 μg/mL, 500 μg/mL and 1000 μg/mL for 12 h, followed by detection of expression of Nampt protein by Western blotting. Microphages were treated with proper concentration of AGEs determined by Western blotting, cells were collected after treatment for 0 h(negative control group), 3 h, 6 h, 12 h and 24 h, and Real-Time PCR and Western blotting were employed to detect the expression of Nampt mRNA and protein in macrophages. Results After exposure to 50 μg/mL,100 μg/mL, 250 μg/mL and 500 μg/mL AGEs,the expression of Nampt protein in macrophages was significantly higher than that in negative control group (P<0.01), and the expression of Nampt protein increased with the concentrations of AGEs, with that after exposure to 250 μg/mL and 500 μg/mL AGEs being higher. Macrophages in the following experiment was treated with 250 μg/mL AGEs. The expression of Nampt mRNA and protein in macrophages treated with 250 μg/mL AGEs was significantly higher than that in negative control group at each time point (P<0.01), and the expression of Nampt mRNA and protein increased with the time of exposure. Conclusion AGEs play a role in the increased expression of Nampt in macrophages, and AGEs may contribute to diabetic atherosclerosis by activating macrophages partly through Nampt pathway.

Key words: advanced glycation end products, macrophage, nicotinamide phosphoribosyltransferase