Akt对IL-1β诱导神经干细胞分化为多巴胺能神经元的作用  

Effect of Akt in the differentiation of dopaminergic neurons induced by IL-1β form the neural stem cells

作　　者：种堔 董美希 刘莹[1] 范国兵 张敏娜[1] 王光辉[1] 吴景龙[2] ZHONG Shen;DONG Meixi;LIU Ying(College of Pharmacy Jining Medical University,Rizhao 276826,China)

机构地区：[1]济宁医学院药学院,山东日照276826 [2]北京理工大学脑科学与神经技术实验室,北京100081

出　　处：《中风与神经疾病杂志》2019年第1期3-6,共4页Journal of Apoplexy and Nervous Diseases

基　　金：国家自然科学基金项目(61473043);国家教育部大学生创新创业基金项目(201610443071)

摘　　要：目的探讨蛋白激酶B/Akt信号途径对IL-1β诱导神经干细胞(neutral stem cells,NSCs)分化多为巴胺能神经元(dopaminergic neurons,DNs)的作用。方法原代取材培养中脑来源大鼠NSCs,用IL-1β诱导NSCs分化;免疫荧光染色法检测βIII-tubulin和酪氨酸羟化酶(TH)表达; Motic Digital Class 1. 1/Motic Images Advanced3. 1图像分析软件测量多巴胺能神经元突起长度; Western blot分析法检测分化细胞Akt磷酸化程度。结果培养细胞表达nestin蛋白,分化为神经元和神经胶质细胞;βIII-tubulin阳性细胞分化率分别为13. 4%±0. 8%(A组)、16. 8%±1. 5%(B组)、28. 8%±2. 4%(C组)、29. 2%±1. 5%(D组)、12. 5%±0. 6%(对照组),βⅢ-tubulin阳性细胞百分率随着IL-1β剂量增加而增加(P <0. 05); TH细胞阳性率分别为1. 6%±0. 5%(A组)、9. 5%±0. 6%(B组)、16. 8%±1. 5%(C组)、16. 6%±1. 2%(D组)、1. 3%±0. 5%(对照组),TH阳性分化细胞随着IL-1β剂量增加而增加(P <0. 01);多巴胺能神经元突起长度分别为(48. 6±8. 3)μm(A组)、(56. 4±5. 2)μm(B组)、(80. 0±12. 8)μm(C组)、(88. 2±10. 5)μm(D组)、(45. 5±6. 5)μm(对照组),多巴胺能神经元突起长度随着IL-1β剂量增加而增加(P <0. 05);经IL-1β诱导后分化细胞Akt的磷酸化水平升高(P <0. 01)。结论 IL-1β作为一种重要的信号分子参与诱导NSCs向DNs分化,分化过程中Akt磷酸化水平升高,蛋白激酶B/Akt信号途径通过磷酸化修饰激活后可能参与NSCs向多巴胺能神经元分化的调节过程。Objective To investigate the effect of Protein kinase B/Akt signal pathway in the process of neural stem cells on the differentiation of dopaminergic neurons induced by IL-1β.Methods The rat neural stem cells form the middle brain were obtained and cultured primitively and the differentiation was induced by IL-1β.Immunofluorescene staining method was used to detect the expression of beta III-tubulin and tyrosine hydroxylase(TH);Motic Digital Class 1.1/Motic Images Advanced 3.1 image analysis software was used to measure neurite’s length of the dopaminergic neurons;Western blot was used to analyze the degree of Akt phosphorylation of differentiated cells induced by IL-1β.Results The cultured cells expressed nestin protein and differentiated into neurons and glial cells;The differentiation rate of beta III-tubulin positive cells was 13.4%±0.8%(group A),16.8%±1.5%(group B),28.8%±2.4%(group C),29.2%±1.5%(group D),12.5%±0.6%(control group),the percentage of beta III-tubulin positive cells increased with the increase of IL-1βdose(P<0.05);The positive rate of TH positive cells was 1.6%±0.5%(group A),6.0%±1.6%(group B),24.8%±1.5%(group C)、24.6%±1.4%(group D)、1.5%±0.5%(control group),TH positive differentiated cells increased with the increase of IL-1βdose(P<0.01);The length of dopaminergic neurons’Bneurites was(48.6±8.3)μm(group A),(56.4±5.2)μm(group B),(80.0±12.5)μm(group C),(80.2±10.5)μm(group D),(45.5±6.5)μm(control group),the neurites’length of dopaminergic neurons increased with the increase of IL-1βdose(P<0.05),and the level of phosphorylation of Akt in differentiated cells increased after IL-1 beta(P<0.01).Conclusion IL-1βinduced neural stem cells differentiate into dopaminergic neurons as a type of crucial signal molecules,the phosphorylation level of Akt increases during the differentiation,and Protein kinase B/Akt signal pathway may be activated by phosphorylation and involved in the regulation of neural stem cells to dopaminergic neurons.

关 键 词：神经干细胞 AKT IL-1Β 多巴胺能神经元 诱导 

分 类 号：R743.3[医药卫生—神经病学与精神病学]