PC12细胞中Vd介导的SPHK1/S1P通路及其在EAE中的潜在功能

目的 探讨维生素D(V_d)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V_d对PC12细胞进行刺激,分为0、6.25、12.5、25、50、100 nmol/L V_d 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V_d刺激PC12细胞24 h后,不同浓度的V_d组SPHK1表达量较0 nmol/L V_d组减少(P<0.05);RT-PCR检测结果显示,不同浓度的V_d组SPHK1较0 nmol/L V_d 组mRNA亦有下调(P<0.05);不同浓度的V_d组S1P合成量较0 nmol/L V_d 组减少(P<0.05)。 结论 V_d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。

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

Objective To investigate the effect of vitamin D(V_d)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 V_d were divided into 6 groups: 0 nmol/L V_d group, 6.25 nmol/L V_d group, 12.5 nmol/L V_d group, 25 nmol/L V_d group, 50 nmol/L V_d group, 100nmol/L V_d 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 V_d for 24h, an obvious change was observed between V_d groups and 0 nmol/L V_d group. Compared with 0nmol/L V_d group, the V_d groups had decreased levels of SPHK1, S1P and SPHK1 mRNA(P<0.05). Conclusion V_d might affect the content of S1P via SPHK1 so as to delay the progression of EAE.