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山东大学学报(医学版) ›› 2016, Vol. 54 ›› Issue (10): 21-24.doi: 10.6040/j.issn.1671-7554.0.2016.224

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PC12细胞中Vd介导的SPHK1/S1P通路及其在EAE中的潜在功能

王震,高计芳,关新媛,郭娟娟,杨笃晓,刘师莲   

  1. 山东大学医学院生物化学与分子生物学系, 山东 济南 250012
  • 收稿日期:2016-03-04 出版日期:2016-10-10 发布日期:2016-10-10
  • 通讯作者: 刘师莲. E-mail: liushilian@sdu.edu.cn E-mail:liushilian@sdu.edu.cn
  • 基金资助:
    国家自然科学基金(81070952,81271319)

Effects of Vd on SPHK1/S1P pathway in PC12 cells and its potential role in EAE

WANG Zhen, GAO Jifang, GUAN Xinyuan, GUO Juanjuan, YANG Duxiao, LIU Shilian   

  1. Department of Biochemistry and Molecular Biology, School of Medicine, Shandong University, Jinan 250012, Shandong, China
  • Received:2016-03-04 Online:2016-10-10 Published:2016-10-10

摘要: 目的 探讨维生素D(Vd)通过鞘氨醇激酶1(SPHK1)影响1-磷酸鞘氨醇(S1P)的合成从而对实验性自身免疫性脑脊髓炎(EAE)发病过程产生的影响。 方法 体外培养PC12细胞,分别用不同浓度的Vd对PC12细胞进行刺激,分为0、6.25、12.5、25、50、100 nmol/L Vd 组。Western blotting检测SPHK1表达量;RT-PCR检测SPHK1 mRNA的表达;ELISA检测S1P合成量。 结果 加入药物Vd刺激PC12细胞24 h后,不同浓度的Vd组SPHK1表达量较0 nmol/L Vd组减少(P<0.05);RT-PCR检测结果显示,不同浓度的Vd组SPHK1较0 nmol/L Vd 组mRNA亦有下调(P<0.05);不同浓度的Vd组S1P合成量较0 nmol/L Vd 组减少(P<0.05)。 结论 Vd可能通过调节SPHK1的表达来影响S1P的合成,以缓解EAE的发病进程。

关键词: 鞘氨醇激酶1, 维生素D, 实验性自身免疫性脑脊髓炎, PC12 细胞, 1-磷酸鞘氨醇

Abstract: Objective To investigate the effect of vitamin D(Vd)on experimental autoimmune encephalomyelitis(EAE)through the regulation of sphingosine kinase type 1(SPHK1)on the content of sphingosine-1-phosphate(S1P). Methods PC12 cells cultured in vitro and stimulated with different concentrations of Vd were divided into 6 groups: 0 nmol/L Vd group, 6.25 nmol/L Vd group, 12.5 nmol/L Vd group, 25 nmol/L Vd group, 50 nmol/L Vd group, 100nmol/L Vd group. The SPHK1 levels were analyzed with Western blotting. The expression of SPHK1 mRNA was detected with RT-PCR. The intracellular S1P was detected with ELISA. Results After PC12 cells were stimulated with different concentrations of Vd for 24h, an obvious change was observed between Vd groups and 0 nmol/L Vd group. Compared with 0nmol/L Vd group, the Vd groups had decreased levels of SPHK1, S1P and SPHK1 mRNA(P<0.05). Conclusion Vd might affect the content of S1P via SPHK1 so as to delay the progression of EAE.

Key words: PC12 cells, Vitamin D, Sphingosine kinase type 1, Sphingosine 1-phosphate, Experimental autoimmune encephalomyelitis

中图分类号: 

  • R744.5+1
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