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摘要：目的 研究维生素D（VD）对大鼠肾上腺嗜铬细胞瘤细胞系PC12细胞中神经酰胺（Cer）水平的影响。方法 采用C2-Cer（20 μmol/L）刺激PC12细胞，MTT法检测Cer对PC12细胞增殖的影响；高效液相色谱串联质谱法（HPLC-MS）测定VD (50 或0 nmol/L)处理的PC12细胞中C16:0-Cer的含量。qRT-PCR法测定VD (50或0 nmol/L)处理的PC12细胞中Cer代谢相关酶mRNA的表达；Western blotting法检测VD（200、100、50、25、12.5和0 nmol/L）处理后，PC12细胞中SPTLC1和Cers6蛋白的相对表达量。结果 C2-Cer能抑制PC12细胞的活性，IC50为2.800 μmol/L。与0 nmol/L VD相比，50 nmol/L VD降低了PC12细胞中C16:0-Cer的含量（19.37±0.61 ng/mL vs 16.50±0.10 ng/mL，P=0.010），升高了SPTLC1 mRNA的表达水平（P=0.001）和Cers6 mRNA的表达水平(P<0.001)，降低了SPTLC1蛋白的相对表达量(P=0.032)，升高了Cers6蛋白的相对表达量(P=0.020)。结论 VD 降低PC12中Cer的含量，可能与降低SPTLC1蛋白表达有关。
Abstract: Objective To study the effect of vitamin D (VD) on the level of ceramide (Cer) in rat adrenal pheochromocytoma cell line PC12. Methods PC12 cells were stimulated with C2-Cer (20 μmol / L) and the effect of Cer on the proliferation of PC12 cells was detected by MTT assay. The inhibitory effect of Cerium on the proliferation of PC12 cells was assayed by HPLC-MS at 50 or 0 nmol / L C16: 0-Cer content in PC12 cells. The mRNA expression of Cer metabolism in PC12 cells treated with VD (50 or 0 nmol / L) was determined by qRT-PCR. The expression of Cer mRNA was detected by Western blotting after VD (200, 100, 50, 25, 12.5 and 0 nmol / L) , Relative expression of SPTLC1 and Cers6 protein in PC12 cells. Results C2-Cer inhibited the activity of PC12 cells with IC50 of 2.800 μmol / L. Compared with 0 nmol / L VD, 50 nmol / L VD reduced C16: 0-Cer content in PC12 cells (19.37 ± 0.61 ng / mL vs 16.50 ± 0.10 ng / mL, P = 0.010) (P = 0.001) and Cers6 mRNA (P <0.001) decreased the relative expression of SPTLC1 protein (P = 0.032) and increased the relative expression of Cers6 protein (P = 0.020). Conclusions Decreasing the content of Cer in PC12 may reduce the expression of SPTLC1 protein.