牡蛎提取物在高温诱导皮质神经干细胞凋亡中的保护作用

背景:课题组前期研究表明,牡蛎提取物对高温所致神经管畸形中的凋亡细胞有一定的保护用用,该提取物对体外培养的神经干细胞的凋亡是否也有保护作用,尚未见报道目的:观察牡蛎提取液在高温所致神经干细胞凋亡中的保护作用.方法:取孕13 d小鼠胚胎大脑皮质细胞培养,采用免疫荧光法检测巢蛋白的表达以鉴定神经干细胞,将培养3代的神经干细胞随机分为4组:高温对照组和牡蛎保护Ⅰ、Ⅱ、Ⅲ组(分别加入质量浓度2.5,5,10 g/L的牡蛎提取液),同时设立培养液对照组(其中无细胞)、培养液+牡蛎提取液对照组(其中无细胞),各牡蛎保护组及对照组均经过39℃高温处理.锥虫蓝染色法检测神经干细胞存活率;MTT法检测神经干细胞的增殖活性;蛋白质印迹方法检测神经干细胞凋亡相关蛋门p53的表达.结果与结论:牡蛎保护Ⅱ、Ⅲ组中细胞的存活率及MTT值明显高于高温对照组(P<0 05);而p53的表达则明显低于高温对照组.提示牡蛎提取液对高温所致神经干细胞的凋亡具有一定的保护作用,这种作用有浓度依赖性.

Protective effect of oyster extract on apoptosis of cerebral neural stem cells induced by hyperthermia

BACKGROUND: Previous results of our study show that oyster extract has some protective effects on apoptosis of the neuroepithelium in neural tube defects induced by hyperthermia in vivo.It remains unclear whether the extraet also protects in vitro cultured neural stem cells.OBJECTIVE: To investigate the protective effect of oyster extract on apoptosis of cerebral neural stem cells induced by hyperthermia.METHODS: The cerebral neural stem cells of embryonic mice of 13 days were cultured in vitro.Nestin expression was detected by immunofluorescence method to identify neural stem cells.The neural stem cells of passage 3 were divided randomly into 4groups: hyperthermia control group and oyster treated Ⅰ,Ⅱ,Ⅲ groups(mass concentration 2.5,5,10 g/L oyster extract solution).In addition,culture solution control group(no cells),and culture solution+oyster extract control group(no cells)were designed.All oyster extract groups and control groups were treated by hyperthermia over 39 ℃.The survival rate and the vitality of neural stem cells were detected by trypan blue staining and MTT assay.Western-blotting was employed to explore the expression of p53 in cerebral neural stem cells of each group.RESULTS AND CONCLUSION: The survival rate and the value of MTT assay in oyster treated groups Ⅱ and Ⅲ were significantly greater than hyperthermia control group(P < 0.05),but the expression of p53 in oyster treated groups Ⅱ and Ⅲ were weaker than hyperthermia control group.Oyster extract plays an important protective role in the apoptosis of neural stem cells induced by hyperthermia.