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-1β dose (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-1β dose (P < 0. 05) , and the level of phosphorylation of Akt in differentiated cells increased after IL-1 beta (P < 0. 01) . Conclusion IL-1β induced 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.%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.
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