Department of Disease Model Animal Research Center, College of Life Science and Technology, Heilongjiang Bayi Agricultural University, Daqing, Heilongjiang 163319, P.R. China.
Pharmaron Beijing Co., Ltd., Beijing 100176, P.R. China.
Mol Med Rep. 2018 Jun;17(6):7827-7834. doi: 10.3892/mmr.2018.8826. Epub 2018 Mar 29.
High concentrations of glutamate may mediate neuronal cell apoptosis by increasing intracellular reactive oxygen species (ROS) levels. Peroxiredoxin V (Prx V), a member of the Prx family, serves crucial roles in protecting cells from oxidative stress. The present study investigated the regulatory effect of Prx V on glutamate‑induced effects on viability and apoptosis in HT22 cells. Western blotting was used for protein expression analysis and Annexin V/PI staining and flow cytometry for determination of apoptosis. The results demonstrated that glutamate may ROS‑dependently increase HT22 cell apoptosis and upregulate Prx V protein levels. Furthermore, knockdown of Prx V protein expression with a lentivirus significantly enhanced HT22 cell apoptosis mediated by glutamate, which was reversed by inhibition of ROS with N‑acetyl‑L‑cysteine. Inhibiting the extracellular signal‑regulated kinase (ERK) signaling pathway with PD98059, a specific inhibitor for ERK phosphorylation, markedly decreased glutamate‑induced HT22 cell apoptosis in Prx V knockdown cells, indicating the potential involvement of ERK signaling in glutamate‑induced HT22 cell apoptosis. In addition, an increase in nuclear apoptosis‑inducing factor was observed in Prx V knockdown HT22 cells following glutamate treatment, compared with mock cells, whereas no differences in B‑cell lymphoma‑2 and cleaved‑caspase‑3 protein expression levels were observed between mock and Prx V knockdown cells. The results of the present study indicated that Prx V may have potential as a therapeutic molecular target for glutamate‑induced neuronal cell death and provide novel insight into the role of Prx V in oxidative‑stress induced neuronal cell death.
高浓度的谷氨酸可能通过增加细胞内活性氧 (ROS) 水平来介导神经元细胞凋亡。过氧化物酶 V (Prx V) 是 Prx 家族的一员，在保护细胞免受氧化应激方面发挥着至关重要的作用。本研究探讨了 Prx V 对谷氨酸诱导的 HT22 细胞活力和凋亡的调节作用。Western blot 用于蛋白质表达分析，Annexin V/PI 染色和流式细胞术用于检测凋亡。结果表明，谷氨酸可能通过 ROS 依赖性增加 HT22 细胞凋亡并上调 Prx V 蛋白水平。此外，用慢病毒敲低 Prx V 蛋白表达显著增强了谷氨酸介导的 HT22 细胞凋亡，而用 N-乙酰-L-半胱氨酸抑制 ROS 则逆转了这种作用。用 ERK 磷酸化的特异性抑制剂 PD98059 抑制细胞外信号调节激酶 (ERK) 信号通路，显著降低了 Prx V 敲低细胞中谷氨酸诱导的 HT22 细胞凋亡，表明 ERK 信号通路可能参与了谷氨酸诱导的 HT22 细胞凋亡。此外，与 mock 细胞相比，在用谷氨酸处理后，Prx V 敲低的 HT22 细胞中观察到核凋亡诱导因子增加，而在 mock 和 Prx V 敲低细胞之间未观察到 B 细胞淋巴瘤-2 和裂解的 caspase-3 蛋白表达水平的差异。本研究结果表明，Prx V 可能作为治疗谷氨酸诱导的神经元细胞死亡的潜在分子靶标，并为 Prx V 在氧化应激诱导的神经元细胞死亡中的作用提供新的见解。
