高等学校化学学报

高等学校化学学报 ›› 2009, Vol. 30 ›› Issue (9): 1759.

• 研究论文 • 上一篇    下一篇

神经鞘磷脂合成酶基因沉默对细胞凋亡的影响

石渊渊, 李志强, 谷敬丽, 王玉兰   

  1. 河南大学医学院, 河南大学神经生物研究所, 开封 475004
  • 收稿日期:2009-03-20 出版日期:2009-09-10 发布日期:2009-09-10
  • 通讯作者: 石渊渊, 女, 副教授, 从事脂代谢与动脉粥样硬化研究. E-mail: syy5161@henu.edu.cn
  • 基金资助:

    国家自然科学基金(批准号: 30670688)、河南省教育厅项目(批准号: 2006180004)和河南大学校内基金(重点理工科)(批准号: 04ZDZR010)资助.

Influence of Sphingomyelin Synthase Knockdown on Cell Apoptosis

SHI Yuan-Yuan*, LI Zhi-Qiang, GU Jing-Li, WANG Yu-Lan   

  1. Medical College of Henan University, Institute of Neurobiology, Henan University, Kaifeng 475004, China
  • Received:2009-03-20 Online:2009-09-10 Published:2009-09-10
  • Contact: SHI Yuan-Yuan. E-mail: syy5161@henu.edu.cn
  • Supported by:

    国家自然科学基金(批准号: 30670688)、河南省教育厅项目(批准号: 2006180004)和河南大学校内基金(重点理工科)(批准号: 04ZDZR010)资助.

PDF (PC)

被引次数

7

摘要：

以HEK293细胞为模型, 利用RNA干扰技术, 将神经鞘磷脂合成酶(SMS)的同工酶(SMS1和SMS2)的siRNA分别联合、转染HEK293细胞. 通过薄层层析法评价SMS酶活性, 同时测神经酰胺(Cer)、卵磷脂(PC)及神经鞘磷脂(SM)的水平, 并以流式细胞仪和Annexin V-FITC、PI双染法检测细胞凋亡. 结果显示, 与对照组相比, SMS基因沉默后SMS表达水平降低(分别降低17%, 20%, 49%); SM水平显著性降低(P<0.05); Cer水平显著性升高(P<0.05); TNF-α诱导的凋亡显著性升高[分别为58%(P<0.01), 24%(P<0.05), 77%(P<0.01)]. 这些结果提示, SMS基因沉默能降低SM水平并升高Cer水平, 明显增加TNF-α诱导的HEK293细胞凋亡. 由于SM是动脉粥样硬化形成的独立危险因子, 因此本研究有可能为动脉粥样硬化的治疗找到新的靶点和有效途径.

关键词: 神经鞘磷脂合成酶; 薄层层析; 神经酰胺; 神经鞘磷脂; 细胞凋亡

Abstract:

HEK293 cells were uesd to investigate the relationship between sphingomyelin synthase(SMS) activity and cell apoptosis. SMS1siRNA and SMS2siRNA were transfected HEK293 cells, respectively, or simultaneously. SMS activity was monitored by thin layer chromatographic assay. Sphingomyelin, ceramide, and phosphatidylcholine levels were determined by an established method. Cell apoptosis was measured by flow cytomertry. The results show that SMS1 and SMS2 knockdown reduces SMS activity(17%, 20%, 49%, respectively), sphingomyelin levels were significantly reduced(P＜0.05), ceramide was significantly increased(P<0.05), compared with control group. We then performed cell apoptosis, and found that SMS1, SMS2, and SMS1/SMS2 knockdown significantly induced by TNF-α mediated apoptosis[58%(P<0.01), 24%(P<0.05), 77%(P<0.01), respectively], suggesting monitoring SMS activity plays an important role on cell apoptosis, thus may have impact on the development of atherosclerosis.

Key words: Sphingomyelin synthase; Thin layer chromatographic; Ceramide; Sphingomyelin; Cells apoptosis

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