Psychiatric Department V, Qingdao Mental Health Center, No. 299, Nanjing Road, Shibei District, Qingdao 266000, China.
Genes (Basel). 2019 Apr 21;10(4):313. doi: 10.3390/genes10040313.
This article explores the mechanism of miR-194 on the proliferation and apoptosis of Aβ-transduced hippocampal neurons. Aβ-transduced hippocampal neuron model was established by inducing hippocampal neurons with Aβ. MTT assay and flow cytometry were used to detect the viability and apoptosis of hippocampal neurons, respectively. qRT-PCR was used to detect changes in miR-194 and Nrn1 expression after Aβ induction. Aβ-transduced hippocampal neurons were transfected with miR-194 mimics and/or Nrn1 overexpression vectors. Their viability and neurite length were detected by MTT assay and immunofluorescence, respectively. Western blot was used to detect protein expression. Aβ inhibited Aβ-transduced hippocampal neuron activity and promoted their apoptosis in a dose-dependent manner. miR-194 was upregulated and Nrn1 was downregulated in Aβ-transduced hippocampal neurons ( < 0.05). Compared with the model group, Aβ-transduced hippocampal neurons of the miR-194 mimic group had much lower activity, average longest neurite length, Nrn1, p-AkT, and Bcl-2 protein expression and had much higher Bax, Caspase-3, and Cleaved Caspase-3 protein expression. Compared with the model group, Aβ-transduced hippocampal neurons of the LV-Nrn1 group had much higher activity, average longest neurite length, Nrn1, p-AkT, and Bcl-2 protein expression and had much lower Bax, Caspase-3, and Cleaved Caspase-3 protein expression. Nrn1 is a target gene of miR-194. miR-194 inhibited apoptosis of Aβ-transduced hippocampal neurons by inhibiting Nrn1 and decreasing PI3K/AkT signaling pathway activity.
本文探讨了 miR-194 对 Aβ 转导海马神经元增殖和凋亡的作用机制。通过诱导海马神经元 Aβ 建立 Aβ 转导海马神经元模型。MTT 法和流式细胞术分别检测海马神经元的活力和凋亡情况。qRT-PCR 检测 Aβ 诱导后 miR-194 和 Nrn1 表达的变化。用 miR-194 模拟物和/或 Nrn1 过表达载体转染 Aβ 转导的海马神经元。用 MTT 法和免疫荧光法分别检测细胞活力和神经突长度。Western blot 法检测蛋白表达。Aβ 呈剂量依赖性抑制 Aβ 转导的海马神经元活性并促进其凋亡。Aβ 转导的海马神经元中 miR-194 上调，Nrn1 下调（ < 0.05）。与模型组相比，miR-194 模拟物组 Aβ 转导的海马神经元活性较低，平均最长神经突长度、Nrn1、p-AkT 和 Bcl-2 蛋白表达较低，Bax、Caspase-3 和 Cleaved Caspase-3 蛋白表达较高。与模型组相比，LV-Nrn1 组 Aβ 转导的海马神经元活性较高，平均最长神经突长度、Nrn1、p-AkT 和 Bcl-2 蛋白表达较高，Bax、Caspase-3 和 Cleaved Caspase-3 蛋白表达较低。Nrn1 是 miR-194 的靶基因。miR-194 通过抑制 Nrn1 降低 PI3K/AkT 信号通路活性，抑制 Aβ 转导的海马神经元凋亡。
