弓形虫抑制神经干细胞向神经元分化的研究  

The effect of excretory-secretory antigens of Toxoplasma gondii on the differentiation of neural stem cells

作　　者：甘小凤[1,2] 张贤[1,2] 程正阳[1,2] 沈继龙[3,2] 罗庆礼[3,2] 余莉[1,2] 

机构地区：[1]安徽医科大学微生物学教研室,安徽合肥230032 [2]人兽共患病安徽省重点实验室 [3]安徽医科大学寄生虫学教研室

出　　处：《中国病原生物学杂志》2014年第11期1004-1008,共5页Journal of Pathogen Biology

基　　金：国家自然科学基金青年基金项目(No.30800965);安徽医科大学青年拔尖人才支持计划项目(No.0113014104)

摘　　要：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 examine the effects of Toxoplasma gondii excretory-secretory antigens（ESA）on the differentiation of neural stem cells. Methods To optimize the differentiation of C17.2neural stem cells,cells were cultured on three different culture media,including DMEM complete medium supplemented with 5% horse serum and 10% fetal bovine serum,serum-free DMEM culture medium,and serum-free DMEM/F12 medium with 2% N2 supplement.The cells were then harvested on day 1,day 3,and day 5to detect levels ofβⅢ-tubulin mRNA using RT-PCR.Afterwards,the cells were further cultured in serum-free DMEM/F12 plus 2% N2 with T.gondii ESA for 5days,and then the harvested cells were analyzed with Western blotting to identify the levels ofβⅢ-tubulin protein. Results The level ofβⅢ-tubulin mRNA in C17.2neural stem cells cultured in DMEM/F12 plus 2% N2supplement（3.93±0.13）was significantly higher than that in cells cultured in DMEM complete medium（0.08±0.34）% and serum-free DMEM culture medium（0）on day 5.When cells were cultured in DMEM/F12 medium with 2% N2 for 5days,the level ofβⅢ-tubulin protein in C17.2neural stem cells was significantly down-regulated as a result of the addition of ESA of T.gondii RH. Conclusion The ESA of RH T.gondii inhibits the differentiation of C17.2neural stem cells into neurons.

关 键 词：弓形虫 神经干细胞 C17.2 分化 

分 类 号：R382.5[医药卫生—医学寄生虫学]