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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SPHK1mRNA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。
Objective To investigate the effect of vitamin D (V_d) on the pathogenesis of experimental autoimmune encephalomyelitis (EAE) through the effect of sphingosine kinase 1 (SPHK1) on the synthesis of sphingosine-1-phosphate (S1P). Methods PC12 cells were cultured in vitro. PC12 cells were stimulated with different concentrations of V_d and divided into 0, 6.25, 12.5, 25, 50 and 100 nmol / L V_d groups. The expression of SPHK1 was detected by Western blotting. The expression of SPHK1 mRNA was detected by RT-PCR and the synthesis of S1P by ELISA. Results Compared with 0 nmol / L V_d group, the expression of SPHK1 in V_d group was significantly decreased (P <0.05) after V_d stimulation of PC12 cells for 24 h. Compared with 0 nmol / L V_d group, L V_d mRNA (P <0.05). Compared with 0 nmol / L V_d group, the amount of S1P in V_d group decreased significantly (P <0.05). Conclusion V_d may affect the synthesis of S1P by regulating the expression of SPHK1 in order to alleviate the pathogenesis of EAE.