山西医科大学学报2024，Vol.55Issue(8)：994-1000,7.DOI:10.13753/j.issn.1007-6611.2024.08.006
1,25(OH)2D3对Aβ1-42诱导阿尔茨海默病细胞模型中细胞焦亡的抑制作用
Inhibitory effect of 1,25(OH)2D3 on Aβ1-42-induced pyroptosis in the cell model of Alzheimer's disease
摘要
Abstract
Objective To investigate the inhibitory effect of 1,25(OH)2D3 on Aβ1-42-induced pyroptosis in the cell model of Alzheimer's disease(AD)and its mechanism.Methods The PC12 cells were divided into control group,model group,Caspase-1-siRNA group,NC-siRNA group,1,25(OH)2D3 group and Caspase-1-siRNA+1,25(OH)2D3 group.The PC12 cells in control group were cultured with DMEM high glucose medium.AD cell model was established with 20 μmol/L Aβ1-42.The PC12 cells in Caspase-1-siRNA group were pretreated with 50 nmol/L Caspase-1-siRNA,and then given 20 μmol/L Aβ1-42.The PC12 cells in NC-siRNA group were treated with 20 μmol/L Aβ1-42 after intervention with 50 nmol/L NC-siRNA.The PC12 cells in 1,25(OH)2D3 group were intervened with 100 nmol/L 1,25(OH)2D3,followed by 20 μmol/L Aβ1-42.The PC12 cells in Caspase-1-siRNA+1,25(OH)2D3 group were firstly pretreated with 50 nmol/L Caspase-1-siRNA,and then added 100 nmol/L 1,25(OH)2D3,followed by 20 μmol/L Aβ1-42 Cellular immunofluorescence was used to observe the expression of apoptosis-associated speck-like protein containing a CARD(ASC).Western blot was used to detect the expressions of pyroptosis-related proteins,including NOD-like receptor thermal protein domain associated protein 3(NLRP3),pro-Caspase-1,cysteinyl aspartate specific proteinase 1(Caspase-1),N-terminal of gasdermin D(GSDMD-N),interleukin-1 β(IL-1β),pro-IL-1β,interleukin-18(IL-18)and pro-IL-18.The cell membrane permeability was detected by acridine orange/ethidine bromide(AO/EB)staining.Results Compared with control group,the fluorescence intensity of ASC protein was enhanced in model group(P＜0.01),the expressions of pyroptosis pathway-related proteins were increased(P＜0.05),and the cell membrane permeability was enhanced(P＜0.01).Compared with model group,the fluorescence intensity of ASC protein was decreased in 1,25(OH)2 D3 group and Caspase-1-siRNA group(P＜0.01),the expressions of pro-Caspase-1,Caspase-1,GSDMD-N,pro-IL-1β,IL-1β,pro-IL-18 and IL-18 were downregulated(P＜0.01),and the cell membrane permeability was reduced(P＜0.01).Compared with Caspase-1-siRNA group,the expressions of GSDMD-N and IL-18 were decreased in Caspase-1-siRNA+1,25(OH)2D3 group(P＜0.01),and the cell membrane permeability was declined(P＜0.01).Conclusion Aβ1-42 could induce the cell pyroptosis in PC12 cells.1,25(OH)2D3 exerts its anti-inflammatory neuroprotective effect in PC12 cells by inhibiting Aβ1-42-induced pyroptosis,and the mechanism is closely related to the inhibition of Caspase-1.关键词
1,25(OH)2D3/Aβ1-42/阿尔茨海默病/细胞焦亡/炎症/Caspase-1Key words
1,25(OH)2D3/Aβ1-42/Alzheimer's disease/pyroptosis/inflammation/Caspase-1分类
医药卫生引用本文复制引用
李学敏,成乐,吕晨慧,王希,陈爽直,张骋,赵海峰..1,25(OH)2D3对Aβ1-42诱导阿尔茨海默病细胞模型中细胞焦亡的抑制作用[J].山西医科大学学报,2024,55(8):994-1000,7.基金项目
中央引导地方科技发展资金资助项目(YDZJSX20231A056) （YDZJSX20231A056）
