首页|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

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

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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。
Mechanism of stilbene glycosides on apoptosis of SH-SY5Y cells via regulating PI3K/AKT signaling pathway
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.

2,3,5,4'-tetrahydroxystilbene 2-O-glucopyranosideAlzheimer diseaseLY294002Phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)Cell proliferationApoptosis

康碧倩、李悦、何晓璇、肖振、胡睿、罗陈亮、乔明宇、吴桂有、李振中、朱晓莹、黄忠仕

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右江民族医学院基础医学院,广西 百色 533000

广西中医药大学药学院,广西 南宁 530200

右江民族医学院药学院,广西 百色 533000

右江民族医学院临床医学院,广西 百色 533000

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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

国家自然科学基金百色市科技计划百色市科技计划右江民族医学院研究生创新计划(2023)

81860709百科20224139百科20211807YXCXJH2023013

2024

海南医学院学报
海南医学院

海南医学院学报

CSTPCD北大核心
影响因子:1.068
ISSN:1007-1237
年,卷(期):2024.30(1)
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