【摘要】 目的 阐明神经细胞特异性表达TRPC1的分子机制，为抑郁症研究提供新的生物学标志。方法 先诱导ESCs在培养基下分化为神经干细胞。通过钙转法用逆转录病毒载体，包装出TRPC1的过表达病毒，感染神经干细胞。再构建出TRPC1基因敲除型胚胎干细胞，诱导分化为TRPC1基因敲除型神经干细胞。获得正常神经干细胞、TRPC1基因敲除型神经干细胞和TRPC1过表达型神经干细胞，对三个细胞系进行加药处理，分别加入10μm地塞米松，分别进行传代增殖数统计和96h胞数计数。结果 TRPC1基因敲除神经干细胞系在正常培养基下其增殖能力会显著减弱；而加药后无论是TRPC1过表达或敲除后的细胞系都对其增殖能力没有显著影响。结论 由此可推论TRPC1或可通过维持神经干细胞的增殖能力起到神经保护作用。更多还原

【Abstract】 Objective To elucidate the molecular mechanism on the specific TRPC1 expression in nerve cells, and provide a novel biological marker for depression research. Methods First, embryonic stem cell(ESCs)was induced to differentiate into neural stem cells in the medium. TRPC1 overexpression virus was packaged with retroviral vector by calcium transfer to infect neural stem cells. Then TRPC1 gene knock-out ESCs were constructed and induced to differentiate into TRPC1gene knock-out neural stem cells. Obtained normal neural stem cells, TRPC1 gene knock-out neural stem cells and TRPC1 over-expressive neural stem cells, three cell lines were treated with the addition 10ug/mL dexamethasone, and the proliferation number and 96h cell number were counted respectively. Results The proliferation ability of TRPC1 gene knock-out neural stem cell line was weakened under the normal culture medium; There was no significant effect on the proliferation of TRPC1 cells either after overexpression or knockout. Conclusions It can be concluded that TRPC1 may play a neuroprotective role by maintaining the proliferation ability of neural stem cells.更多还原