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目的探讨弓形虫排泄分泌抗原(ESA)对神经干细胞分化的影响。方法分别用含5%马血清、10%胎牛血清DMEM完全培养液,无血清DMEM培养液及含2%N2的DMEM/F12培养液培养诱导神经干细胞系C17.2分化,分别于诱导分化后1、3、5d收集细胞,提取RNA,采用RT-PCR检测神经元标志蛋白βⅢ-tubulin基因的转录;用含ESA的2%N2DMEM/F12培养液培养C17.2细胞5d,收集细胞,提取总蛋白,采用Western blot检测βⅢ-tubulin蛋白水平。结果用含2%N2的DMEM/F12培养液培养5d,C17.2细胞βⅢ-tubulin基因转录水平为3.93±0.13,另两组分别为(0.08±0.34)%和0,差异有统计学意义(P<0.05);2%N2诱导分化后5d,弓形虫ESA能显著降低βⅢ-tubulin蛋白水平(P<0.05)。结论弓形虫ESA能抑制C17.2神经干细胞向神经元分化。
Objective To investigate the effect of Toxoplasma gondii secreting antigen (ESA) on the differentiation of neural stem cells. Methods The neural stem cell line C17.2 was induced by culture in DMEM medium containing 5% horse serum, 10% fetal bovine serum, serum-free DMEM and DMEM / F12 containing 2% N2, respectively. The cells were harvested at day 1, day 3, day 5 and RNA was extracted. Transcription of the neuronal marker βⅢ-tubulin gene was detected by RT-PCR. C17.2 cells were cultured with 2% N2DMEM / F12 medium containing ESA for 5 days. Protein, using Western blot detection of β Ⅲ-tubulin protein levels. Results The transcript level of βⅢ-tubulin in C17.2 cells was 3.93 ± 0.13 in DMEM / F12 culture medium containing 2% N2 for 5 days, and the difference was statistically significant (0.08 ± 0.34)% and 0 P <0.05). 5 days after 2% N2 induction, ESA of Toxoplasma gondii decreased the level of βⅢ-tubulin (P <0.05). Conclusion Toxoplasma gondii ESA can inhibit the differentiation of C17.2 neural stem cells into neurons.