摘要
目的探讨参麦微球及其在Wnt信号通路下对骨髓间质干细胞(BMSC)分化为神经元细胞的诱导作用。方法分离BMSC并传代培养后进行细胞鉴定。取第3代BMSC,分为空白组及诱导组,分别采用含空白微球及参麦微球的无血清培养基进行诱导,3 h后检测两组细胞的神经丝蛋白-M(NF-M)、神经元特异性烯醇化酶(NSE)、巢蛋白、微管联合蛋白-2(MAP-2)、生长相关蛋白-43(GAP-43)、胶质纤维酸性蛋白(GFAP)的表达。将第3代BMSC分为Wnt3a、Wnt5a组及对照组,分别采用诱导液1(含Wnt3a及参麦液微球)、诱导液2(含Wnt5a及参麦液微球)、单纯诱导剂(只含参麦液微球)进行培养。培养2周后,检测3组细胞巢蛋白、NSE和GFAP的mRNA水平。结果诱导组NF-M、巢蛋白、GAP-43的平均吸光度值以及NSE、MAP-2阳性率均高于空白组(均P <0. 05)。Wnt3a组的NSE和GFAP mRNA相对表达水平均高于Wnt5a组及对照组(均P <0. 05),而Wnt5a组的NSE和GFAP mRNA相对表达水平与对照组比较差异均无统计学意义(均P> 0. 05)。结论参麦微球可诱导BMSC向神经元细胞方向分化,而Wnt3a信号通路对其有促进作用。
Objective To explore Shenmai microspheres and its induction effect under Wnt signaling pathway on differentiation of bone marrow mesenchymal stem cell(BMSC) into neuronal cell. Methods BMSCs were identified after isolation and subculture.The third-generation BMSCs were divided into blank group and induction group,receiving induction by serum-free medium containing blank microspheres and serum-free medium containing Shenmai microspheres,respectively,3 hours later,expression of neurofilament-M(NF-M),neuron-specific enolase(NSE),nestin, microtubule -associated protein-2(MAP-2),growth-associated protein-43(GAP-43) and glial fibrillary acidic protein(GFAP) was detected in both groups.The third-generation BMSCs were divided into Wnt3a group,Wnt5a group and control group,cultured using induction solution 1(containing Wnt3a and Shenmai microspheres), induction solution 2(containing Wnt5a and Shenmai microspheres),and simple inducer(containing Shenmai microspheres alone), respectively .The levels of nestin,NSE and GFAP mRNAs were measured in the three groups after 2 weeks of culture. Results The mean absorbance values of NF-M,nestin and GAP-43,as well as the positive rates of NSE and MAP-2,in the induction groups were higher than those in the blank group(all P <0.05).The relative expression levels of NSE and GFAP mRNAs in the Wnt3a group were higher than those in the Wnt5a group and control group(all P <0.05),but there was no statistically significant difference between the Wnt5a group and the control group in relative expression level of NSE or GFAP mRNA(all P >0.05). Conclusion Shenmai microspheres can induce differentiation of BMSC into neuronal cell,which might be promoted by Wnt3a signaling pathway.
作者
李志彬
郭玉海
邢庆嘉
刘权锋
LI Zhi-bin;GUO Yu-hai;XING Qing-jia;LIU Quan-feng(The Second Clinical College,Guangzhou University of Chinese Medicine,Guangzhou 510000,China)
出处
《广西医学》
CAS
2019年第8期999-1003,共5页
Guangxi Medical Journal
基金
国家自然科学基金(81273780)
关键词
骨髓间质干细胞
神经元细胞
细胞分化
WNT信号通路
微球
参麦
中药
Bone marrow mesenchymal stem cell
Neuronal cell
Wnt signaling pathway
Microspheres
Shenmai
Chinese medicine
