川陈皮素调节YAP/TAZ信号通路对OGD/R诱导神经元细胞铁死亡的影响  

Nobiletin protects OGD/R-induced neuronal cell ferroptosis by regulating the YAP/TAZ signaling pathway

作　　者：张玲 高子红 潘斌鲜 ZHANG Ling;GAO Zihong;PAN Binxian(Department of Neurology,Dachuan District People’s Hospital(Dazhou Third People’s Hospital),Sichuan,Dazhou 635000,China)

机构地区：[1]四川省达州市达川区人民医院(四川省达州市第三人民医院)神经内科,635000

出　　处：《河北医药》2024年第21期3222-3226,共5页Hebei Medical Journal

基　　金：四川省卫生健康委员会科研课题(编号:19PJ016)。

摘　　要：目的探究川陈皮素(NOB)调节Yes相关蛋白(YAP)/转录共激活因子PDZ结合基序(TAZ)信号通路对氧糖剥夺/复糖复氧(OGD/R)诱导的神经元铁死亡的影响。方法常规培养小鼠海马神经元HT22细胞,采用OGD/R诱导HT22建立细胞损伤模型,将HT22细胞随机分为对照组(Control组)、模型组(OGD/R组)、低浓度NOB组(NOB-L组,15μmol/L NOB)、高浓度NOB组(NOB-H组,30μmol/L NOB)和高浓度NOB+YAP激活剂XMU-MP-1组(NOB-H+XMU-MP-1组,30μmol/L NOB+5μmol/L XMU-MP-1)。CCK-8法检测5组HT22细胞活力;酶联免疫吸附实验(ELISA)检测5组HT22细胞谷胱甘肽(GSH)、丙二醛(MDA)水平;活性氧(ROS)试剂盒检测细胞ROS水平。特异性荧光探针法检测细胞内铁含量;Western Blot检测5组HT22细胞中YAP/TAZ信号通路相关蛋白和铁死亡指标蛋白溶质载体家族7成员11(SLC7A11)、谷胱甘肽过氧化物酶4(GPX4)、酰基合成酶长链家族成员4(ACSL4)的表达。结果与Control组比较,OGD/R组HT22细胞OD_(450)(48 h、72 h)、GSH水平、SLC7A11、GPX4蛋白表达显著下降(P<0.05),MDA、ROS水平、铁离子水平、ACSL4、YAP、TAZ蛋白表达显著上升(P<0.05)。与OGD/R组比较,NOB-L组、NOB-H组HT22细胞相关指标变化与上述相反(P<0.05)。YAP激活剂XMU-MP-1减轻了NOB对OGD/R诱导的神经元铁死亡的抑制作用。结论NOB可能通过抑制YAP/TAZ信号通路减轻OGD/R诱导的神经元铁死亡。Objective To investigate the effect of Nobiletin(NOB)on neuronal ferroptosis induced by oxygen glucose deprivation/glucose reoxygenation(OGD/R)by regulating the Yes-associated protein(YAP)/transcriptional coactivator with PDZ-binding domain(TAZ)signaling pathway.Methods HT22 cells from mouse hippocampal neurons were cultured routinely,and a cell injury model was established by inducing OGD/R in HT22 cells.HT22 cells were induced with blank control,OGD/R,low-dose NOB(15μmol/L NOB),high-dose NOB(30μmol/L NOB)and high-dose NOB+YAP inhibitor XMU-MP-1(30μmol/L NOB+5μmol/L XMU-MP-1).CCK-8 assay was applied to detect the activity of HT22 cells in each group.Enzyme linked immunosorbent assay(ELISA)was applied to detect the levels of glutathione(GSH)and malondialdehyde(MDA)in HT22 cells of each group.Reactive oxygen species(ROS)assay kit was used to detect cellular ROS level.Specific fluorescence probe method was applied to detect intracellular iron content.Western blot was applied to detect the protein expressions of YAP/TAZ signaling pathway-related proteins and iron death marker protein solute carrier family 7 member 11(SLC7A11),glutathione peroxidase 4(GPX4),and acyl coenzyme A synthetase long chain family member 4(ACSL4)in HT22 cells in each group.Results Compared with those of blank control,OGD/R-induced HT22 cells presented significantly lower optical density at 450nm(OD450)at 48h and 72h,GSH level and protein levels of SLC7A11 and GPX4,but higher MDA and ROS levels,iron ion level and protein levels of ACSL4,YAP and TAZ(P<0.05).Compared with those of OGD/R-induced cells,HT22 cells induced with low-dose and high-dose NOB showed opposite changes in the above indicators(P<0.05).Treatment of YAP inhibitor XMU-MP-1 alleviated NOB-induced inhibitory effect on OGD/R-induced ferroptosis in neurons.Conclusion NOB may alleviate OGD/R-induced neuronal ferroptosis by inhibiting the YAP/TAZ signaling pathway.

关 键 词：川陈皮素 Yes相关蛋白/转录共激活因子PDZ结合基序信号通路 氧糖剥夺/复糖复氧 铁死亡 

分 类 号：R743.31[医药卫生—神经病学与精神病学]