天津医药

天津医药 ›› 2017, Vol. 45 ›› Issue (3): 254-257.doi: 10.11958/20161043

• 实验研究 • 上一篇    下一篇

ADMA 对内皮生长晕细胞凋亡及 JNK 表达的影响

张福青， 李新△， 胡亚会， 焦占全， 田晓琳， 刘杰   

  1. 天津医科大学第二医院神经内科 （邮编 300211）
  • 收稿日期:2016-09-23 修回日期:2017-02-24 出版日期:2017-03-15 发布日期:2017-03-21
  • 基金资助:
    天津市卫计委科技基金 （2014KZ107）

Effects of asymmetric dimethylarginine on apoptosis and expression of c-Jun N-terminal kinase in endothelial outgrowth cells

ZHANG Fu-qing, LI Xin△, HU Ya-hui, JIAO Zhan-quan, TIAN Xiao-lin, LIU Jie   

  1. Department of Neurology, the Second Hospital of Tianjin Medical University, Tianjin 300211, China
  • Received:2016-09-23 Revised:2017-02-24 Published:2017-03-15 Online:2017-03-21

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摘要： 摘要： 目的 探讨非对称二甲基精氨酸 （ADMA） 对内皮生长晕细胞 （EOCs） 凋亡的影响及凋亡相关 c-Jun 氨基末端激酶（JNK）表达水平的变化。方法 从脐带血中分离培养 EOCs 并进行鉴定， 与不同浓度 ADMA（0、 1、 5、 10、 20µmol/L）共同培育 48 h。在 10 µmol/L ADMA 培养液中加入不同浓度（0、 5、 10、 20、 40 µmol/L）JNK 特异性抑制剂SP600125， 流式细胞仪检测细胞凋亡率， Western blot 检测 Caspase-3 及磷酸化 JNK（p-JNK）水平。结果 1、 5、 10、20 µmol/L ADMA 可诱导 EOCs 凋亡， 浓度越高凋亡率越高 （F=5.681， P＜0.05）。ADMA 还可提高凋亡因子 Caspase-3 的水平（F=6.733， P＜0.05）， 诱导 JNK 磷酸化。SP600125 可缓解 ADMA 造成的 EOCs 细胞凋亡， 抑制 Caspase-3及 p-JNK 的表达。结论 ADMA 可诱导 EOCs 细胞凋亡， 此作用可能通过激活 JNK 信号转导途径实现。

关键词: 精氨酸, 细胞凋亡, 蛋白激酶类, JNK 丝裂原活化蛋白激酶类, 非对称性二甲基精氨酸, 内皮生长晕细胞

Abstract: Abstract： Objective To investigate the effects of asymmetric dimethylarginine (ADMA) on apoptosis and phosphorylation of c-jun N-terminal kinase (JNK) in endothelial outgrowth cells (EOCs). Methods The mononuclear cells were harvested from umbilical cord blood by ficoll density gradient centrifugation, and induced into EOCs and then expanded in vitro. The identified EOCs were treated with different concentrations of ADMA (0, 1, 5, 10, 20 µmol/L) for 48 h.The adherent cells were treated with 10 µmol/L ADMA， then different concentrations of JNK specific inhibitor SP600125 (0，5， 10， 20 and 40 µmol /L) were added and incubated for 48 hours. Caspase-3 activity was measured by microplate reader.Apoptotic incidences of EOCs were quantitatively determined by flow cytometry. The expression of Caspase- 3 and phosphorylase- JNK (p- JNK) were detected by Western blot assay. Results The treatment of ADMA (1- 20 µmol/L)significantly induced apoptosis in EOCs by enhancing Caspase- 3 express and also induced phosphorylation of JNK (P＜0.05). Meantime, the JNK specific inhibitor SP600125 could attenuate the apoptosis induced by ADMA during this process(F=6.733， P＜0.05) and inhibit the expression of Caspase- 3 and p- JNK. Conclusion ADMA can induce apoptosis in EOC, which may be achieved by activating JNK signal transduction pathway.

Key words: arginine, apoptosis, protein kinases, JNK mitogen-activated protein kinases, asyrmetric dimethylarginine, endothelial outgrowth cells