Aβ25-35 Induces Abnormal Expression of Per2 by Calcium Overload in HT22 Nerve Cell

Li Yameng; Zhao Xueling; Ren Yanan; Zhang Rui; Zhao Jin; Yuan Yuan; Wang Li; Wang Xiaohui

doi:10.16289/j.cnki.1002-0837.2017.06.006

Abstract：Objective To investigate the effect of calcium overload on the abnormal expression of Per2 induced by amyloid-beta protein 25-35 (Aβ25-35) in HT22 nerve cell.Methods In vivo study,the circadian rhythm of C57BL/6 mice was observed by the mice wheel-running experiment after bilateral hippocampal injection of Aβ25-35 (300 μmoL/kg) and sterile water.In vitro study,HT22 cells were divided into control group,Aβ25-35 treatment group and Nim + Aβ25-35 treatment group.Cell viability was assessed with MTT assay.The amount of per2 mRNA at different CT times were quantified by utilizing real-time fluorescence quantitative PCR,and the expression of PER2 protein at CT16 were evaluated by Western Blotting and immunofluorescence staining.The intracellular calcium concentration was detected by flow cytometry.Results A325-35 in-duced circadian rhythm disorder in C57BL/6 mice.Compared with control group,5,10,15,20,25,30 μmoL/L A325-35 decreased the cell viability significantly (P ＜ 0.05).Compared with control group,20 μmol/L Aβ25-35 decreased the per2 mRNA levels and the expression of PER2 protein respectively significantly at CT16(P ＜ 0.05).The calcium antagonist nimodipine can antagonize the toxicity of Aβ25-35 by increasing the per2 mRNA levels and the expression of PER2 protein significantly at CT16(P ＜ 0.05).Conclusion Calcium overload plays an important role in the abnormal expression of Per2 induced by Aβ25-35 in HT22 nerve cell.