二苯乙烯苷调控PI3K/AKT信号通路干预SH-SY5Y细胞凋亡的机制研究
Mechanism of stilbene glycosides on apoptosis of SH-SY5Y cells via regulating PI3K/AKT signaling pathway
康碧倩 1李悦 1何晓璇 2肖振 1胡睿 1罗陈亮 2乔明宇 1吴桂有 2李振中 3朱晓莹 4黄忠仕1
作者信息
- 1. 右江民族医学院基础医学院,广西 百色 533000
- 2. 广西中医药大学药学院,广西 南宁 530200
- 3. 右江民族医学院药学院,广西 百色 533000
- 4. 右江民族医学院临床医学院,广西 百色 533000
- 折叠
摘要
目的:探讨二苯乙烯苷(TSG)经PI3K/AKT通路对冈田酸诱导的人神经母细胞瘤细胞(SH-SY5Y)凋亡的影响及调控机制.方法:CCK-8法筛选OA最佳浓度,将SH-SY5Y细胞分为对照组、模型组、TSG组、LY294002组和LY294002+TSG组.通过CCK-8法和TUNEL法检测各组细胞增殖和凋亡情况;Western blotting法和qRT-PCR法检测PI3K、P-PI3K(Y607)、AKT、P-AKT(Ser473)、Bcl-2、Bax蛋白及mRNA表达情况,通路及凋亡蛋白相对表达量以P-PI3K(Y607)/PI3K、P-AKT(Ser473)/AKT、Bcl-2/Bax灰度比值表示.结果:OA最佳处理浓度为40 nmol/L.与对照组相比,模型组细胞活力降低,细胞凋亡率升高,通路及凋亡蛋白、PI3K、AKT、Bcl-2 mRNA表达水平降低,Bax mRNA表达水平升高(均P<0.05);与模型组相比,TSG组细胞活力升高,细胞凋亡率降低,通路及凋亡蛋白、PI3K、AKT、Bcl-2 mRNA相对表达水平升高,Bax mRNA表达水平降低(均P<0.05),LY294002组细胞活力降低,细胞凋亡率升高,P-PI3K(Y607)/PI3K蛋白表达水平显著降低(P<0.05)、P-AKT(Ser473)/AKT、Bcl-2/Bax蛋白表达水平较明显降低,但未有统计学意义,PI3K、AKT、Bcl-2 mRNA表达水平降低,Bax mRNA表达水平升高(均P<0.05);与LY294002组相比,LY294002+TSG组细胞活力升高,细胞凋亡率降低,通路及凋亡蛋白相对表达水平、PI3K、AKT、Bcl-2 mRNA升高,Bax mRNA表达水平降低(均P<0.05).结论:二苯乙烯苷可能通过干预PI3K/AKT信号通路,进而调节Bcl-2、Bax等凋亡因子的表达,从而缓解冈田酸诱导的SH-SY5Y细胞凋亡.
Abstract
Objective:To investigate the effects of stilbene glycoside(TSG)on okadaic acid-induced apoptosis in human neuroblastoma cells(SH-SY5Y)via the PI3K/AKT pathway.Methods:The optimal concentration of OA was screened by CCK-8 assay,and SH-SY5Y cells were divided into the control group,the model group,the TSG group,the LY294002 group and the LY294002+TSG group.The proliferation and apoptosis in each group were detected by CCK-8 and TUNEL assays;Western blotting method and real-time fluorescence quantitative polymerase chain reaction were used to detect the expression of PI3K,P-PI3K(Y607),AKT,P-AKT(Ser473),Bcl-2 and Bax proteins.The relative protein expression was represented by P-PI3K(Y607)/PI3K,P-AKT(Ser473)/AKT and Bcl-2/Bax gray ratio.Results:CCK-8 screened the optimal concentration of OA as 40 nmol/L.Compared with the control group,the model group increased relative cell viability,decreased apoptosis rate,the pathway and apoptotic proteins expression levels of P-PI3K(Y607)/PI3K,P-AKT(Ser473)/AKT and Bcl-2/Bax were de-creased,the mRNA expression levels of PI3K,AKT and Bcl-2 were decreased,While Bax mRNA expression level increased(P<0.05);Compared with model group,the TSG group increased relative cell viability,decreased apoptosis rate,increased protein expression levels of P-PI3K(Y607)/PI3K,P-AKT(Ser473)/AKT,Bcl-2/Bax,and increased mRNA expression levels of PI3K,AKT,and Bcl-2,While Bax mRNA expression decreased(P<0.05),the LY294002 group decreased relative cell viabili-ty,increased apoptosis rate,P-PI3K(Y607)/PI3K protein expression levels were significantly decreased(P<0.05),P-AKT(Ser473)/AKT and Bcl-2/Bax protein expression levels were significantly decreased,but there was no statistical significance.PI3K,AKT and Bcl-2 mRNA expression levels were decreased,and Bax mRNA expression levels were increased(all P<0.05);Compared with the LY294002 group,the LY294002+TSG group increased relative cell viability,decreased apoptosis rate,and the protein expression levels of P-PI3K(Y607)/PI3K,P-AKT(Ser473)/AKT and Bcl-2/Bax were increased.The mRNA expression levels of PI3K,AKT,Bcl-2 were increased,and Bax was decreased(all P<0.05).Conclusion:Stilbene gly-coside may alleviate okadaic acid-induced apoptosis in SH-SY5Y cells by interfering with the PI3K/AKT signaling pathway,which in turn regulates the expression of apoptotic factors such as Bcl-2 and Bax.
关键词
二苯乙烯苷/阿尔茨海默症/磷酸肌醇-3激酶抑制剂/磷酸肌醇3-激酶/蛋白激酶B/细胞增殖/细胞凋亡Key words
2,3,5,4'-tetrahydroxystilbene 2-O-glucopyranoside/Alzheimer disease/LY294002/Phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)/Cell proliferation/Apoptosis引用本文复制引用
基金项目
国家自然科学基金(81860709)
百色市科技计划(百科20224139)
百色市科技计划(百科20211807)
右江民族医学院研究生创新计划(2023)(YXCXJH2023013)
出版年
2024
国家科技期刊平台
NSTL
维普
万方数据