目的 探讨氯化锂对缺氧后神经干细胞(NSCs)磷脂酰肌醇-3-羟激酶/蛋白激酶B(PI3K/Akt)信号通道活性的影响。方法 分离培养新生SD乳鼠(SD)NSCs,建立缺氧NSCs模型。正常对照组NSCs加无血清培养基,单纯缺氧组仅缺氧,生理盐水干预组缺氧后加生理盐水,三组氯化锂干预组分别加入氯化锂至所需的浓度。免疫组织化学技术检测各组Akt /P-Akt阳性细胞表达。结果 缺氧NSCs形态不规则,胞体肿胀,甚至出现细胞膜破裂,数量减少,折光性减弱;缺氧NSCs死亡数较未缺氧NSCs明显增多、活性明显降低(P均<0.05)。1 mM氯化锂干预组P-Akt表达较单纯缺氧组、生理盐水干预组及5 mM氯化锂干预组强,较3 mM氯化锂干预组弱。3 mM 氯化锂干预组P-Akt表达较1 mM 、5 mM氯化锂干预组强。3 mM氯化锂干预组P-Akt阳性细胞数较单纯缺氧组、生理盐水干预组、1 mM、5 mM氯化锂干预组明显增多(P<0.05)。结论 氯化锂激活缺氧后NSCs PI3K/Akt信号通路。
Abstract
Objective To investigate the effect of lithium chloride on PI3K/Akt signaling pathway in hypoxic neural stem cells (NSCs). Methods The models of hypoxic NSCs were established by culturing and identify NSCs isolated from neonatal SD rat.The NSCs of normal control group was cultured in serum-free medium.The culture medium of normal saline intervention group was immediately added normal saline,and lithium chloride intervention group was immediately added different concentration of lithium chloride.Akt /P-Akt signaling pathway markers of NSCs were detected by immunohistochemistry. Results The amount of NSCs decreased and the characteristics of the NSCs was abnormal in shape,such as low refraction,swollen perikarya or even membranolysis.The number of dead cells of NSCs in hypoxic group increased significantly (P<0.05).The activeness of NSCs in hypoxic group decreased significantly (P<0.05).Compared with hypoxic group,normal saline intervention group and 5 mM lithium chloride intervention group,P-Akt expression level of hypoxic NSCs in 1 mM lithium chloride intervention group increased significantly,while decreased significantly compared with 3 mM lithium chloride intervention group.Compared with 1 mM and 5 mM lithium chloride intervention group,P-Akt expression level of hypoxic NSCs in 3 mM lithium chloride intervention group increased significantly.Compared with other groups,the number of hypoxic NSCs positive expressing P-Akt in 3 mM lithium chloride group increased significantly (P<0.05). Conclusions Lithium chloride activates PI3K/Akt signaling pathway in hypoxic NSCs.
关键词
神经干细胞 /
缺氧模型 /
信号通路 /
氯化锂
Key words
neural stem cells /
hypoxia model /
signaling pathway /
lithium chloride
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基金
中国博士后科学基金(20070410505)
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