人参环氧炔醇对体外培养RSC96细胞的实验观察

目的研究人参环氧炔醇(Panaxydol,PND)对体外培养RSC96细胞神经营养因子及髓鞘蛋白表达的影响并探讨其机制。方法用含不同血清浓度的培养基培养RSC96细胞,MTT检测其增殖能力,以获取最适宜RSC96细胞体外生长的血清浓度。在适宜的血清培养基中加入PND(10μmol/L)处理RSC96细胞,以RT-PCR、western blot及ELISA检测RSC96细胞NGF和BDNF的表达。另外,预先在培养基中加入钙离子通道阻滞剂尼非地平探讨PND可能的作用途径。结果培养基血清浓度为4mmol/L时,RSC96细胞生长形态最接近于原代Schwann细胞。PND增强RSC96细胞表达和释放NGF和BDNF(P0.05)。尼非地平的使用,则削弱了PND对RSC96细胞的作用(P0.05)。结论在适宜的血清培养基中,体外培养的RSC96细胞可替代原代Schwann细胞,作为研究药物对它的影响及机制的细胞模型。PND增强RSC96细胞的生物活性的作用机制可能通过Ca2+信号途径介导。

The effects of Panaxydol on the cultured RSC96 cell lines in vitro

Objective　To investigate the effects of Panaxydol (PND) on the expression of NGF and myelin protein in cultured RSC96 cells.　Methods　RSC96 cells were cultured and decided the optimal culture condition by adjusting the serum concentration. After detecting the proliferation of RSC96 cells by MTT assay, the cultures were added with PND (10 μmol/L), or without PND as the control. After 24 h, the expression and secretion of NGF and BDNF in cultured cells were examined by RT-PCR, western blot and ELISA. For exploring the possible intracellular signals involved in PND effects, N-type Ca2+ channel blocker - Nifedipine was co-cultured for 4 h before the addition of PND.　Results　The result showed that, 4 mmol/L serum in the culture medium was the best concentration for the growth of RSC96 cells. PND significantly enhanced the expression and release of NGF and BDNF of RSC96 cells. However, the effects of PND on the RSC96 cells can be partly eliminated by Nifedipine.　Conclusions　The results indicate that RSC96 cells cultured in an optimal serum medium could replace primary Schwann cells as the subject for in vitro researches, furthermore, the effect of PND on the RSC96 cells is mediated partly via Ca2+ signal.