S31-201经下调p-STAT3与Notch1诱导神经干细胞向神经元分化  

S31-201 promotes neuronal differentiation of neural stem cells via downregulation of p-STAT3 and Notch

作　　者：崔猛[1,2] 冯世庆[1] 范宁建[1] 

机构地区：[1]天津医科大学总医院骨科,天津市300052 [2]天津医科大学骨科临床学院,天津市300211

出　　处：《中国矫形外科杂志》2013年第14期1435-1441,共7页Orthopedic Journal of China

基　　金：国家自然科学基金资助项目(编号:81070982)

摘　　要：[目的]观察抑制信号传导和转录激活子3(signal transducer and activator of transcription 3,STAT3)信号通路对神经干细胞(neuronal stem cells,NSCs)的分化及Notch1表达的影响,进一步明确STAT3在调控NSCs分化过程中的作用及与Notch1的协同关系。[方法]从孕14～15 d的SD大鼠胚胎大脑皮质中分离NSCs,体外培养传代。STAT3抑制剂为S31-201。分组:对照组与三个不同浓度的S31-201组(2.5μmol/L、5μmol/L、10μmol/L)。体外培养48 h后,用Western blotting方法观察S31-201对NSCs中p-STAT3蛋白表达的影响;体外培养48 h与72 h后,用细胞免疫荧光化学染色方法观察S31-201对NSCs中微管相关蛋白2(microtubule-associated protein2,MAP-2)、胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)表达的影响;体外培养48 h与72 h后,用RT-PCR与Western blotting方法观察S31-201对NSCs中Notch1基因与蛋白表达的影响。[结果]三个浓度的S31-201组均观察到NSCs中p-STAT3表达的下调,以10μmol/L浓度组抑制作用最强(P<0.05)。S31-201在48 h与72h两个时间点均能促进NSCs中MAP-2表达增加并抑制NSCs中GFAP的表达(P<0.05)。S31-201在48与72 h两个时间点均能抑制NSCs中Notch1 mRNA与蛋白的表达(P<0.05)。[结论]S31-201可以有效抑制NSCs中p-STAT3的水平;抑制p-STAT3的表达能促进NSCs向神经元分化并抑制NSCs向星形胶质细胞分化;抑制p-STAT3可下调Noteh1的表达,二者可能协同调控NSCs的分化。[ Objective] To study the effects of inhibition of the Stat3 signaling pathway on neural stem cell（NSC） differen- tiation and Notchlexpression, and to further clarify the function of Star3 and the relationship between Stat3 and Notchl in the regulation of NSC differentiation. [ Method] NSCs were isolated from the embryonic cerebral cortex of 14 - 15 days pregnant SD ratsand then cultured and passaged in vitro. Third -generation NSCs were used for the experiment. S31 -201 was used as the Stat3 inhibitor. Cells were divided into a control group called the natural differentiation group and 3 groups that were exposed to different doses of S31 -201 （2.5 μM, 5 μM, and 10 μM）. After 48 h culture in vitro,the effect of S31 -201 on phospho- Stat3 protein expression in NSCs was observed by western blotting. Furthermore, after 48 h and 72 h culture in vitro, the impact of S31 -201 on MAP- 2 and Gfap expression in NSCs was observed by immunohistochemical analyses, and the effect on NotchlmRNA and protein expression in NSCs was observed using RT - PCR and western blotting. [ Result] p - STAT3 levels in NSCs were downregulated in the 3 S31 - 201 groups; the strongest inhibition occurred withl 0 μM of the inhibitor （ P 〈 0.05 ）. S31 -201 promotedMap-2 expressionand inhibited Gfap expression in NSCs after 48 h and 72 h culture in vitro（P 〈 0.05 ）. S31 - 201 also suppressed Notchl mRNA and protein expression in NSCs at these time points （P 〈 0.05）. [ Conclusion] S31 - 201 can effectively inhibit the level of p- STAT3 in NSCs,inhibition of p- STAT3 can promote NSCs to differentiate into neu- rons and inhibit their differentiation into astrocytes. Moreover, p -STAT3 inhibition can decrease the expression of Notchl. Our findings suggest that Stat3 and Notchl regulate the differentiation of NSCs cooperatively.

关 键 词：NSCS 信号转导和转录激活子3 NOTCH1 S31-201 分化 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]