Cognitive Impairment
Increased glycogen synthase kinase 3β activity involves in the decreased expression of Bmal1 induced by amyloid-beta protein 31-35 in HT22 cells
Guo Shuai, Sun Cong, Wang Changtu, Yuan Yuan, Ning Na, Hou Xiaohong, Wang Li, Wang Xiaohui
Published 2020-02-08
Cite as Chin J Neurol, 2020,53(02): 96-102. DOI: 10.3760/cma.j.issn.1006-7876.2020.02.004
Abstract
ObjectiveTo investigate the effect of glycogen synthase kinase 3β (GSK3β) on the decreased expression of Bmal1 induced by amyloid-beta protein 31-35 (Aβ31-35) in HT22 cells.
MethodsHT22 mouse hippocampal cells were divided into control group, Aβ31-35 group and LiCl+Aβ 31-35 group by random number table method in the present study. Cells were synchronized to G0/G1 phase by 1% serum starvation for 1 hour (circadian time 0 (CT0)). Cell viability was detected by the cell counting kit-8 assay. The mRNA expression of clock gene Bmal1 was examined by real-time PCR at different CT times. The expression of GSK3β and BMAL1 protein was detected by Western blotting.
ResultsCompared with the control group, Aβ31-35 induced the decreased expression of Bmal1 mRNA; The expression of both Bmal1 mRNA and BMAL1 protein was decreased significantly at CT20 (Bmal1 mRNA: 0.38±0.06 vs 0.83±0.08, t=4.549, P=0.001; BMAL1 protein: 0.67±0.04 vs 1.00±0.04, t=5.943, P<0.001). In the Aβ31-35 group, GSK3β activity was increased and the ratio of phosphorylated GSK3βS9 to GSK3β was decreased compared to the control group (0.66±0.08 vs 1.02±0.14, t=2.217, P=0.025). Aβ31-35 decreased the viability of HT22 cells (71.85%±6.20% in the Aβ31-35 group vs 98.14%±2.68% in the control group, t=3.891, P=0.006), and the GSK3β inhibitor LiCl pretreatment effectively reversed the decline of the viability induced by Aβ31-35 (90.74%±5.74% in the LiCl+Aβ31-35 group vs 71.85%±6.20% in the Aβ31-35 group, t=3.412, P=0.010). LiCl (in the LiCl+Aβ31-35 group) increased the expression of Bmal1 mRNA and BMAL1 protein significantly at CT20 compared with the Aβ31-35 group (Bmal1 mRNA: 0.72±0.05 vs 0.38±0.06, t=4.378, P=0.001; BMAL1 protein: 0.90±0.04 vs 0.67±0.04, t=4.052, P=0.002).
ConclusionIncreased GSK3β activity involved in the decreased expression of Bmal1 induced by Aβ31-35 in HT22 cells.
Key words:
Amyloid; Glycogen synthase kinase 3; Bmal1 gene; HT22 cells
Contributor Information
Guo Shuai
Department of Pathology, Shanxi Medical University, Taiyuan 030001, China
Sun Cong
Department of Pathology, Shanxi Medical University, Taiyuan 030001, China
Wang Changtu
Department of Pathology, Shanxi Medical University, Taiyuan 030001, China
Yuan Yuan
Department of Morphology, Shanxi Medical University, Taiyuan 030001, China
Ning Na
Department of Pathology, Shanxi Medical University, Taiyuan 030001, China
Hou Xiaohong
Department of Pathology, Shanxi Medical University, Taiyuan 030001, China
Wang Li
Department of Pathology, Shanxi Medical University, Taiyuan 030001, China
Wang Xiaohui
Department of Pathology, Shanxi Medical University, Taiyuan 030001, China
