姜黄素通过激活cAMP-PKA-CREB信号通路促进诱导多功能干细胞神经分化  

Curcumin Promotes Neural Differentiation of Induced Pluripotent Stem Cells via CAMP-PKA-CREB Pathway

作　　者：姚辉[1] 舒涛[1] 徐义春[1] 侯刚[1] 赵慧清[1] 张俊斌[1] YAO Hui;SHU Tao;XU Yi-chun;HOU Gang;ZHAO Hui-qing;ZHANG Jun-bin(Department of Orthopaedics,The Third Affiliated Hospital of Sun Yat-sen University,Guangzhou,510630)

机构地区：[1]中山大学附属第三医院骨科,广州510630

出　　处：《中国中西医结合杂志》2021年第5期602-607,共6页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：广东省中医药局科研项目(No.20191071)。

摘　　要：目的探讨姜黄素促诱导多功能干细胞(iPSCs)定向神经分化的作用及机制。方法将分化的神经干细胞(NSCs)及神经元细胞分为空白组、姜黄素组及姜黄素+H89组,其中空白组采用维甲酸(RA)诱导培养,姜黄素组在上述基础上加入1μmol/L姜黄素,姜黄素+H89组加入1μmol/L姜黄素和20μmol/L H89(PKA抑制剂)。采用免疫荧光染色观察各组NSCs及神经元细胞的分化情况,ELISA法测定各组cAMP含量,Western Blot法测定各组PKAc、pCREB、CREB、Nestin及MAP2蛋白表达。结果免疫荧光染色显示iPSCs经RA诱导可分化为NSCs并进一步分化为神经元细胞。在分化的NSCs及神经元细胞中,姜黄素组及姜黄素+H89组cAMP含量较空白组增加(P<0.05);NSCs中姜黄素组Nestin、PKAc、pCREB表达较空白组均增加(P<0.05);姜黄素+H89组Nestin、PKAc、pCREB表达较空白组及姜黄素组均减少(P<0.05)。神经元细胞中,姜黄素组MAP2、PKAc、pCREB表达较空白组均增加(P<0.05);姜黄素+H89组MAP2、PKAc、pCREB表达较姜黄素组均减少(P<0.05)。结论姜黄素可以促进i PSCs分化为NSCs,并进一步分化为神经元细胞,而这一促进效应可能是通过激活cAMP-PKA-CREB信号通路实现。Objective To explore the effect and mechanism underlying curcumin promotes neural differentiation of induced pluripotent stem cells(i PSCs).Methods IPSC-derived neural stem cells(NSCs)and neurons were divided into three groups,ie,blank group,curcumin group and curcumin+H89 group.The iPSCs were induced by retinoic acid(RA)in the blank group,while by 1μmol/L curcumin added in the curcumin group and 1μmol/L curcumin+20μmol/L H89(inhibitor of PKA)added in the curcumin+H89 group.Immunofluorescence staining was used to observe the differentiation of NSCs and neurons in each group.CAMP content in each group was evaluated by ELISA.Protein expression of PKAc,p CREB,CREB,Nestin and MAP2 were assessed by Western Blot.Results Immunofluorescence staining showed iPSCs were induced into NSCs and neurons by RA.The cAMP content of the differentiated NSCs and neurons in curcumin group and curcumin+H89 group were both higher than that in blank group(P<0.05).The expressions of Nestin,PKAc,and pCREB in NSCs in curcumin group were higher than those in blank group(P<0.05),and the corresponding expressions in curcumin+H89 group were significantly lower than those in both blank group and curcumin group(P<0.05).The expressions of MAP2,PKAc and pCREB in neuros in curcumin group were higher than those in blank group(P<0.05),and the corresponding expressions in curcumin+H89 group were lower than those in curcumin group(P<0.05).Conclusion It is suggested that curcumin promotes neural differentiation of iPSCs through the activation of cAMP-PKA-CREB pathway.

关 键 词：姜黄素 诱导多功能干细胞 神经分化 cAMP-PKA-CREB信号通路 

分 类 号：R285.5[医药卫生—中药学]