上海交通大学学报（医学版）

›› 2011, Vol. 31 ›› Issue (5): 533-.doi: 10.3969/j.issn.1674-8115.2011.05.002

• 论著（基础研究） • 上一篇    下一篇

高糖和甲基乙二醛对人脐静脉内皮细胞Ang-Tie2信号链表达的影响

徐雅虹, 方 炜, 袁江姿, 戴慧莉, 倪兆慧, 钱家麒   

  1. 上海交通大学 医学院附属仁济医院肾脏科 |上海市腹膜透析研究中心, 上海 200127
  • 出版日期:2011-05-28 发布日期:2011-05-27
  • 通讯作者: 方 炜, 电子信箱: fangwei_sh@126.com。
  • 作者简介:徐雅虹(1985—), 女, 硕士；电子信箱: spot_dog@126.com。
  • 基金资助:

    国家自然科学基金 (30600290)和上海市科委基金（044119620, 07QA14040, 08dz1900501）

Effects of high glucose and methylglyoxal on expression of angiopoietin 2 in cultured human umbilical vascular endothelial cells

XU Ya-hong, FANG Wei, YUAN Jiang-zi, DAI Hui-li, NI Zhao-hui, QIAN Jia-qi   

  1. Renal Division, Renji Hospital, Shanghai Jiaotong University School of Medicine, Shanghai Center for Peritoneal Dialysis Research, Shanghai 200127, China
  • Online:2011-05-28 Published:2011-05-27
  • Supported by:

    National Natural Science Foundation of China, 30600290；Shanghai Science and Technology Committee Foundation, 044119620, 07QA14040, 08dz1900501

PDF (PC)

1931

摘要：

目的 观察高浓度葡萄糖对体外培养的人脐静脉内皮细胞（HUVECs）血管生成素2（Ang2）和酪氨酸激酶2受体（Tie2）表达的影响，探讨腹膜透析患者腹膜血管新生的可能机制。方法 分别以4.25%葡萄糖（高糖组）、4.25%甘露醇（甘露醇组）和800 μmol/L葡萄糖降解产物甲基乙二醛（MGO）（MGO组）对体外培养的HUVECs进行干预，设立空白对照组。以MTT比色法测定的D(490 nm)比较各组HUVECs干预后12、24、48 h时点的细胞增殖情况；分别采用Real-Time PCR技术、ELISA法和Western blotting法检测各组HUVECs中Ang2和Tie2 mRNA表达、细胞培养上清液中Ang2分泌量及HUVECs中Tie2蛋白的表达。结果 高糖组、甘露醇组和 MGO组HUVECs干预后12 h时点的D(490 nm)均明显高于空白对照组（P<0.05），干预后24 h时点的D(490 nm)则显著低于空白对照组（P<0.05）。与空白对照组比较，高糖组、甘露醇组和MGO组HUVECs中Ang2 mRNA表达均明显上调，高糖组和甘露醇组细胞培养上清液中Ang2分泌量均显著增加（P<0.05），MGO组细胞培养上清液中Ang2分泌量则明显减少（P<0.05）。高糖组、甘露醇组和MGO组HUVECs中Tie2 mRNA和蛋白表达均显著低于空白对照组（P<0.05）。结论 透析液中的高浓度葡萄糖可能通过刺激内皮细胞增殖和上调Ang2表达而促进腹膜血管新生；葡萄糖具有独立于其高渗透压之外的促血管新生作用。

关键词: 血管新生, 血管生成素, 酪氨酸激酶2受体, 腹膜透析

Abstract:

Objective To investigate the effects of high glucose on expression of angiopoietin 2 (Ang2) and tyrosine kinase receptor (Tie2) in human umbilical vascular endothelial cells （HUVECs）cultured in vitro, and explore the possible mechanism of peritoneal angiogenesis in patients with peritoneal dialysis. Methods HUVECs cultured in vitro were treated with 4.25% glucose (high glucose group), 4.25% mannitol (mannitol group) and 800 μmol/L methylglyoxal (MGO) (MGO group), and blank control group was establishment. D(490 nm) was measured by MTT assay to compared the cell proliferation of HUVECs after incubation for 12 h, 24 h and 48 h in each group. The expression of Ang2 and Tie2 mRNA in HUVECs, the secretory volume of Ang2 in supernatant of cell culture and the expression of Tie2 protein in HUVECs were detected by RealTime PCR, ELISA and Western blotting respectively in each group. Results D(490 nm) of HUVECs in high glucose group, mannitol group and MGO group were significantly higher than that in blank control group after treatment for 12 h （P<0.05）, and D(490 nm) of HUVECs in high glucose group, mannitol group and MGO group were significantly lower than that in blank control group after treatment for 24 h （P<0.05）. The expression of Ang2 mRNA in HUVECs in high glucose group, mannitol group and MGO group was significantly higher than that in blank control group （P<0.05）, the secretory volumes of Ang2 in supernatant of cell culture in high glucose group and mannitol group were significantly higher than that in blank control group （P<0.05）, and the secretory volume of Ang2 in supernatant of cell culture in MGO group was significantly lower than that in blank control group （P<0.05）. The expression of Tie2 mRNA and protein in HUVECs in high glucose group, mannitol group and MGO group was significantly lower than that in blank control group （P<0.05）. Conclusion High glucose may promote angiogenesis in peritoneum through stimulating the endothelial cell proliferation and regulating the expression of Ang2.

Key words: angiogenesis, angiopoietin, tyrosine kinase receptor, peritoneal dialysis