天津医药

• 细胞与分子生物学 • 上一篇    下一篇

PI3K抑制剂对人系膜细胞增殖的影响

马莉1,张春江1,李宏2,赵丹2,陈建平3,杨军2,杨晓萍3   

  1. 1. 石河子大学医学院
    2. 新疆石河子大学医学院第一附属医院
    3. 石河子大学医学院第一附属医院肾病科
  • 收稿日期:2014-05-07 修回日期:2014-06-25 出版日期:2014-10-15 发布日期:2014-10-15
  • 通讯作者: 杨晓萍

Effect of PI3K Inhibitor on Proliferation of Human Glomerular Mesangial Cells

  • Received:2014-05-07 Revised:2014-06-25 Published:2014-10-15 Online:2014-10-15

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摘要：

【摘要】目的  探讨磷脂酰肌醇3 激酶（PI3K）抑制剂LY294002 对体外培养的人肾小球系膜细胞增殖的影响，为进一步研究人系膜细胞增殖的发生机制提供依据。方法 体外培养人系膜细胞，同步化后，用含有0.02、0.2、2、 20、100 mg/L 浓度的LY294002 进行干预处理，倒置相差显微镜动态观察细胞的生长，用细胞增殖与活性检测试剂盒 CCK-8 检测人系膜细胞的增殖情况；选择抑制人系膜细胞增殖作用较佳的LY294002 浓度, 分别培养0.5、1、24 、48 h 后用CCK-8 试剂盒检测细胞的增殖情况。结果 0.02 mg/L LY294002 对人系膜细胞无明显的抑制作用（P＞0.05），抑制率仅为5.05%；在0.2~20 mg/L 浓度范围内，LY294002 抑制细胞增殖的作用随浓度的增大而增强，呈剂量依赖性（P＜0.05），当浓度大于20 mg/L 时，抑制率可达96.75%，细胞停止生长。2 mg/L LY294002 作用0.5 h 对人系膜细胞增殖的抑制不明显（P＞0.05）；作用1 h 具有显著的抑制作用（P＜0.05）；作用1~48 h，LY294002 抑制细胞增殖的能力逐渐增强（P＜0.05）。结论 LY294002 在一定浓度和时间范围内具有抑制人系膜细胞增殖的作用，PI3K/Akt/ mTOR 信号通路可能调控人系膜细胞的增殖。

关键词: 1-磷脂酰肌醇3-激酶, 肾小球系膜细胞, PI3K/Akt/mTOR信号通路, 细胞增殖, LY294002

Abstract:

[Abstract] Objective  To investigate the effect of LY294002 on proliferation of cultured human glomerular mesangial cells, and to study on the mechanism underlying this proliferation. Methods  Different concentrations of LY294002 (0.02，0.2，2，20，100 mg/L) were administrated to the cultured human glomerular mesangial cells. The effects of mesangial cell proliferation were measured using CCK-8 colorimetric assay. Under appropriate concentrations, proliferation of cultured mesangial cells were measured using CCK-8 at 0.5, 1, 24, 48 hours of drug administration time. Results  LY294002（0.02 mg/L）didn’t obviously inhibited the growth of mesangial cells（P＞0.05）, the inhibition rate was 5.05%. The effect of LY294002 on the cells decreased with rising concentration (0.2 to 20 mg/L) in a dose-dependent manner (P<0.05). Mesangial cells would stop growing with the increasing concentration. LY294002 didn’t obviously inhibited the growth of mesangial cells at 2 mg/L during 0 to 1 h（P＞0.05）, but inhibited proliferation gradually from 1 to 48 h (P < 0.05). Conclusion  Within the certain range of concentration and time, the LY294002 inhibits the proliferation of human glomerular mesangial cells, and PI3K/Akt/mTOR signaling pathway may regulate the proliferation of human mesangial cell.

Key words: 1-phosphatidylinositol 3-kinase, mesangial cells, PI3-Kinase-Akt-mTOR pathway, cell proliferation, LY294002