镉对小鼠胚胎神经干细胞向胶质细胞分化的影响

Influence of cadmium chloride on differentiation of neural stem cells into glial cells

目的:探讨氯化镉对小鼠胚胎神经干细胞向胶质细胞分化的影响。方法:以0、0.1、0.3、1.0、3.0、10.0μmol/L的氯化镉对小鼠胚胎神经干细胞染毒,通过四噻氮唑盐(MTT)法测定染毒24和48 h后的细胞毒性;用神经干细胞核转录因子(Sox2)和胶原纤维酸性蛋白(GFAP)分别鉴定神经干细胞和星形胶质细胞,采用细胞免疫荧光分析小鼠神经干细胞向星形胶质细胞的分化情况。结果:与空白对照组相比,随氯化镉染毒剂量的增加,细胞存活率逐渐降低,呈剂量依赖性下降(24 h,r=-0.91;48 h,r=-0.96;P均<0.05),10.0μmol/L作用24 h组以及1.0、3.0和10.0μmol/L作用48 h组与空白对照组的细胞存活率相比,差异均有统计学意义(P<0.05或P<0.01)。分化48 h后,Sox2阳性细胞率仅在10.0μmol/L出现下降(P<0.05);而GFAP阳性细胞百分率随着剂量升高均减少,且呈剂量-效应关系(r=-0.99,P<0.05)。结论:氯化镉可以抑制神经干细胞向星形胶质细胞的分化。

OBJECTIVE：To investigate the effects of cadmium chloride on differentiation of mouse neural stem cells（mNSCs） in vitro.METHODS：Primary culture of mNSCs was used as research model.mNSCs were exposed to cadmium chloride（0,0.1,0.3,1.0,3.0 and 10.0μmol/L）.MTT assay was used to access cytotoxicity at 24 and 48 h,the differentiation of mNSCs after 48 h exposure of cadmium chloride was determined by immunocytochemistry staining. RESULTS：With the increase of cadmium chloride dose,cytotoxicity on mNSCs was accentuated in a dose-dependent manner（24 h,r= -0.91;48 h,r= -0.96;P0.05）.The cell viability at 10.0μmol/L after 24 hours and at 1.0, 3.0 and 10.0μmol/L after 48 h were lower than the control（P0.05 or P0.01）.Cadmium chloride influenced differentiation of mNSCs.10.0μmol/L significantly decreased the rate of Sox2-positive cells（P 0.05） after 48 h exposure to cadmium chloride.The GFAP-positive also decreased in a dose-dependent manner（r = - 0.99,P 0.05）. CONCLUSION：Cadmium chloride could inhibit the differentiation of mNSCs to astrocytes.