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目的：研究不同强度电刺激对脂肪干细胞（adipose tissue-derived stem cells, ADSC）向神经元方向分化的作用。方法：在细胞电刺激室内对ADSCs 进行电刺激(10Hz, 1h)，强度选用五个强度，分别为0 V/cm、0.5 V/cm、1.0 V/cm、3.0 V/cm、5.0 V/cm；电刺激后，对细胞进行流式凋亡检测及CCK-8 活性检测，明确不同电刺激对ADSCs 的影响；同时，应用免疫荧光染色、Western Blotting 评估各组细胞神经特异性标志物microtubule-associated protein 2 (MAP-2)与beta-tubulin 的表达情况。应用RT-PCR 检测MAP-2、 beta-tubulin 和neurofilaments 200(NF-200)的mRNA 水平，评价不同强度电刺激对ADSCs 其向神经方向分化的影响。结果：1V/cm 的电刺激，未引起明显的细胞凋亡，同时促进了细胞增殖。此外，1V/cm电刺激后，细胞中的MAP-2 和beta-tubulin 的免疫荧光染色强度及蛋白含量显著提高；MAP-2、beta-tubulin 和NF-200 的mRNA及蛋白量显著提高。3V/cm 及更高强度的电刺激可导致凋亡细胞数目显著增加。结论：强度为1V/cm 的电刺激可促进脂肪干细胞的增殖，并促进其向神经方向分化，为神经损伤的治疗提供了新的可行方法。

英文摘要:

Objective:To explore the role of electrical stimulation (ES) in neuronal differentiation of adipose tissue-derived stem cells (ADSCs).Methods:Cultured ADSCs were exposed to ES (10Hz, 1h) at different intensities (0 V/cm, 0.5 V/cm, 1.0 V/cm, 3.0 V/cm, 5.0 V/cm). The proliferation of cells was characterized by using a CCK-8 assay. The apoptosis of cells after ES was examined by flow cytometry. The expression of MAP-2 and beta-tubulin were detected by immunofluorescence staining and Western blotting, and the mRNA levels of MAP-2，beta-tubulin and NF-200 were measured by RT-PCR to examine neuronal differentiation of ADSCs.Results:ES at 1V/cm was shown to promote cell proliferation without increasing the rate of apoptosis. In addition, the immunofluorescence intensity and protein levels of MAP-2 and beta-tubulin were significantly increased compared to shamstimulated cells. Further studies showed that the mRNA levels of MAP-2, beta-tubulin and NF-200 were significantly increased by ES at 1 V/cm, while ES at 3V/cm or higher intensities resulted in more cell apoptosis.Conclusion:ES at 1 V/cm is able to promote the neuronal differentiation of ADSCs and is beneficial for the proliferation of ADSCs, which highlights the potential application in the treatment of neurodegenerative diseases and neurotrauma in nervous system.

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