论文部分内容阅读
目的:丙戊酸盐(丙戊酸valproic acid,VPA)在过去数十年应用于临床治疗癫痫和偏头痛。然而,母亲怀孕早期使用VPA将大大增加子代罹患孤独症群谱障碍的易感性。鉴于Wnt/β-catenin信号通路对神经元增殖、分化、突起生长及凋亡的重要作用,本文旨在研究VPA在原代培养神经元中对Wnt/β-catenin信号通路的影响。方法:用VPA处理原代培养神经元,以生理盐水处理为对照,运用Western Blot检测Wnt/β-catenin通路相关信号分子Wnt1,Wnt2,WIF-1,Dickkopf 1及效应分子β-catenin的表达变化,同时运用免疫荧光技术观察神经元形态变化。结果:与对照组比较,VPA处理显著增加Wnt1及Wnt2的表达(P<0.05~0.01),而未增加WIF-1及Dickkopf 1的表达(P>0.05);VPA处理也导致Wnt/β-catenin通路活性上调,表现为神经元内β-catenin含量显著上升。此外,与对照组比较,VPA处理促进神经元生长,表现为神经元突起数目(P<0.05~0.01)及总长度显著增加(P<0.05~0.01);Wnt/β-catenin通路抑制剂能部分抑制VPA引起的Wnt/β-catenin通路活性上调及神经元生长。结论:VPA通过上调Wnt/β-catenin信号通路促进神经元生长,可能是VPA增加子代罹患孤独症群谱障碍易感性的原因。
Purpose: Valproic acid (VPA) has been used clinically for the treatment of epilepsy and migraine for decades. However, the use of VPA early in pregnancy in maternal mothers will greatly increase the susceptibility of offspring to spectrum disorders. In view of the important role of Wnt / β-catenin signaling pathway in proliferation, differentiation, neurite outgrowth and apoptosis, we aimed to investigate the effect of VPA on Wnt / β-catenin signaling in primary cultured neurons. Methods: Primary cultured neurons were treated with VPA, and normal saline was used as a control. The expression of Wnt1, Wnt2, WIF-1, Dickkopf 1 and β-catenin in Wnt / β-catenin pathway were detected by Western Blot At the same time, immunofluorescence was used to observe the morphological changes of neurons. Results: Compared with the control group, VPA treatment significantly increased the expression of Wnt1 and Wnt2 (P <0.05 ~ 0.01), but did not increase the expression of WIF-1 and Dickkopf 1 (P> 0.05) Pathway activity is increased, showing a significant increase in β-catenin content in neurons. In addition, compared with the control group, VPA treatment promoted the neuron growth, showing the number of neurons (P <0.05-0.01) and the total length significantly increased (P <0.05-0.01); Wnt / β-catenin pathway inhibitor Inhibit VPA induced Wnt / |Â-catenin pathway activity and neuronal growth. Conclusion: VPA can promote neuron growth by up-regulating the Wnt / β-catenin signaling pathway, which may be the reason VPA increases susceptibility to autism spectrum disorder in offspring.