摘要
目的探讨FCGBP在Aβ_(25-35)诱导的SH-SY5Y细胞凋亡和炎症中的作用。方法选取GEO数据库中的GSE3300数据集分析并筛选阿尔茨海默病(AD)组织中的差异表达基因。用0、5、10和20μmol/L的淀粉样β蛋白25-35(Aβ_(25-35))诱导SH-SY5Y细胞24 h后,用Western blotting法检测FCGBP的表达变化,以明确构建体外AD细胞模型的最佳Aβ_(25-35)浓度。随后将si-FCGBP及其阴性对照si-NC转染至SH-SY5Y细胞中,用20μmol/L Aβ_(25-35)处理24 h后,用Western blotting评估细胞转染效率,CCK-8检测细胞活力,Annexin V-FITC/PI法和Western blotting检测细胞凋亡和凋亡相关蛋白的表达,ELISA检测细胞上清中炎症因子的水平。结果分析GSE3300数据集发现FCGBP在AD组织中的表达量较正常组织明显升高(P<0.05)。20μmol/L Aβ_(25-35)处理SH-SY5Y细胞24 h为最佳AD细胞建模条件。敲低FCGBP可促进Aβ_(25-35)诱导的SH-SY5Y细胞的活力(P<0.05),减少细胞的凋亡率(P<0.05),上调Bcl-2(P<0.05),下调Bax和cleaved Caspase3(P均<0.05),抑制炎症因子IL-1β、IL-6、IL-18和TNF-α的表达(P均<0.05)。结论FCGBP在AD脑组织中高表达,敲低FCGBP可以促进Aβ_(25-35)诱导的SH-SY5Y细胞的活力、抑制细胞凋亡、减少炎症因子的释放。
Objective To investigate the role of FCGBP in Aβ_(25-35)-induced apoptosis and inflammation in SH-SY5Y cells.Methods The GSE3300 data set in GEO database was selected to analyze and screen the differentially expressed molecules in Alzheimer's disease(AD)tissues.SH-SY5Y cells were induced with 0,5,10 and 20μmol/L amyloid-βpro⁃tein 25-35(Aβ_(25-35))for 24 h,and the expression of FCGBP was detected by Western blotting to determine the optimal con⁃centration of Aβ_(25-35)for constructing AD cell model in vitro.Subsequently,si-FCGBP and its negative control si-NC were transfected into SH-SY5Y cells,and treated with 20μmol/L Aβ_(25-35)for 24 h.The transfection efficiency was evaluated by Western blotting.The cell viability was detected by CCK-8.Annexin V-FITC/PI and Western blotting were used to detect apoptosis and apoptosis-related protein expression.ELISA was used to detect the inflammatory factors in cell supernatant.Results Analysis of GSE3300 data set showed that the expression of FCGBP in AD tissues was significantly higher than that in normal tissues(P<0.05).Treatment of SH-SY5Y cells with 20μmol/L Aβ_(25-35)for 24 h was the best condition for AD cell modeling.Knockdown of FCGBP promoted the viability of SH-SY5Y cells induced by Aβ_(25-35)reduced the apoptosis rate of cells,up-regulated Bcl-2,down-regulated Bax and cleaved Caspase3,and inhibited the expression of inflammatory factors IL-1,IL-6,IL-18 and TNF-α(all P<0.05).Conclusions FCGBP is highly expressed in AD brain tissue.Knockdown of FCGBP promotes the viability of SH-SY5Y cells induced by Aβ_(25-35),inhibits apoptosis and reduces the re⁃lease of inflammatory factors.
作者
董帅
靳晓华
杨立军
陈丽
李向华
DONG Shuai;JIN Xiaohua;YANG Lijun;CHEN Li;LI Xianghua(Department of Geriatrics,Zhangqiu District People's Hospital,Jinan 250200,China;Department of Neurology,Zhangqiu District People's Hospital,Jinan 250200,China)
出处
《老年医学研究》
2025年第3期59-63,共5页
Geriatrics Research
