Wnt/β-catenin通路在脉冲强磁场促神经干细胞增殖中的作用初探

Role of Wnt/β-catenin pathway in proliferation of neural stem cells induced by high-intensity pulsed electromagnetic fields

目的:摸索抑制神经干细胞(NSCs)中β-catenin表达的合适条件,为获得抑制β-catenin表达的NSCs做准备。利用抑制β-catenin表达的NSCs,探索wnt/β-catenin通路在脉冲强磁场促NSCs增殖中的作用。方法:取新生的SD大鼠双侧侧脑室下的NSCs,然后用无血清培养基培养14d。利用RNAi原理构建β-catenin shRNA慢病毒载体及阴性慢病毒载体,并转染NSCs,确定合适的感染复数(MOI)值。将NSCs分为阴性组(阴性病毒转染组)和实验组(β-catenin shRNA病毒转染组),并用4T、0.1Hz、8次的脉冲强磁场干预。在用脉冲强磁场干预后的1d、7d,采用CCK8法检测阴性组和实验组的NSCs增殖情况。结果:β-catenin shRNA序列在MOI=10时,对NSCs中β-catenin的抑制效果最好。阴性病毒组在脉冲强磁场干预后1d、7d时NSCs增殖最明显(P<0.05),而实验组在干预前后NSCs增殖水平无明显变化。结论:wnt/β-catenin通路在脉冲强磁场促NSCs增殖中起作用。

Objective：To investigate the optimal conditions of inhibiting theβ-catenin expression in neural stem cells（NSCs）for harvesting NSCs low-expressingβ-catenin,and exploring the roles of Wnt/β-catenin pathway in proliferation of NSCs induced by the high-intensity pulsed electromagnetic fields.Methods：（1）NSCs were derived from subventricular zone（SVZ）of newborn SD rats,and then cultured for 14 days using serum-free medium.（2）The RNAi method was used to inhibitβ-catenin expression in NSCs,and the optimal MOI for transfection to inhibitβ-catenin levels in NSCs was measured.（3）The NSCs were divided into the negative control group and experimental group,and then they were exposed in the high-intensity pulsed electromagnetic fields with stimulus condition of 4 T,0.1 Hz,8 times.（4）At 1 st and 7 th day after the intervention with the high-intensity pulsed electromagnetic fields,the proliferation of NSCs was tested by the CCK8 assay.Results：（1）When MOI=10,the best inhibitory effects onβ-catenin could be obtained in NSCs.（2）At 1 st and 7 th day after NSCs were exposed to the high-intensity pulsed electromagnetic fields,the proliferation of NSCs in the negative control group was significantly higher than in the experimental group（P〈0.05）.Conclusion：The Wnt/β-catenin pathway may play a role in proliferation of NSCs induced by the high-intensity pulsed electromagnetic fields.