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作 者:史文涛 戴瑶 毕士奇 赵有发 张志坚[2] Shi Wentao;Dai Yao;Bi Shiqi;Zhao Youfa;Zhang Zhijian(Nanjing Gaochun People's Hospital,Jiangsu University,Nanjing,Jiangsu 211302;School of Medicine Zhenjiang,212001 China)
机构地区:[1]南京市高淳人民医院,南京211302 [2]江苏大学医学院,镇江212001
出 处:《神经解剖学杂志》2018年第5期567-571,共5页Chinese Journal of Neuroanatomy
基 金:国家自然科学基金(81571830)
摘 要:目的:研究氯倍他索(Clo)对神经干细胞自发分化的影响,为Clo治疗中枢性神经损伤提供医学理论基础。方法:利用无血清悬浮法分离培养及鉴定大鼠NSCs;选取第三代NSCs,以含10%胎牛血清完全培养基贴壁诱导分化7 d,培养基中各加入同体积二甲基亚砜、2. 5、5μmol/L的Clo;免疫荧光染色和Western Blot方法检测NSCs分化后轴突膜蛋白(GAP-43)、酸性胶质蛋白(GFAP)、髓磷脂碱性蛋白(MBP)表达情况,进一步探讨Clo的作用机制。结果:分离培养的NSCs高表达Nestin; Clo作用组表达GAP-43和MBP较对照组明显提高(P <0. 05),GFAP表达水平低于对照组(P <0. 05),Clo作用组表达音猬因子(SHH)和锌指蛋白(Gli 1)均高于对照组(P <0.05)。结论:Clo可在体外通过SHH信号通路促进NSCs向神经元细胞和少突胶质细胞分化,与此同时Clo可抑制NSCs向星形胶质细胞分化。Objective: To study the effects of clobetasol( Clo) on the spontaneous differentiation of rat neural stem cells,provide a theoretical basis for Clo to treat central nerve injury.Methods: The serum-free suspension method was used to isolate and culture the rat NSCs.The third-generation NSCs were selected and adhered to the 10% FBS serumcontaining medium to induce differentiation for 7 days.Clobetasol 0( Same volume of Dimethyl sulfoxide),2.5 and 5μmol/L were added to the medium.Immunofluorescent staining and Western Blot methods were used to detect the expression of growth associated protein-43( GAP-43),glial fibrillary acidic protein( GFAP) and myelin basic protein( MBP)after differentiation of NSCs,and Nestin was used to identify NSCs.Results: Nestin was highly expressed in NSCs.The expression of GAP-43 and MBP in Clo treated group was significantly higher than those in control group( P〈0.05); the expression level of GFAP was lower than that in control group( P〈0.05) and the expression of sonic hedeahog( SHH)and Gli 1 were higher than those of control group( P〈0.05).Conclusion: Clo could promote the differentiation of NSCs into neurons and oligodendrocytes through the SHH signal pathway in vitro,and meanwhile while inhibit the differentiation of NSCs into astrocytes.
关 键 词:NSCS 氯倍他索 神经元细胞 星形胶质细胞 少突胶质细胞
分 类 号:R338[医药卫生—人体生理学]
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