白藜芦醇通过IRE1α-XBP1通路抑制衣霉素诱导的神经元凋亡和GSK-3β/Tau蛋白磷酸化  

Resveratrol inhibits tunicamycin-induced neuronal apoptosis and phosphorylation of GSK-3β/Tau protein through IRE1α-XBP1 pathway

作　　者：林淼 刘芸如 于佳欣 张文轩 劳凤学[1] 黄汉昌[1,2] LIN Miao;LIU Yun-ru;YU Jia-xin;ZHANG Wen-xuan;LAO Feng-xue;HUANG Han-chang(Institute of Functional Factors and Brain Sciences,Beijing Union University;Key Laboratory of Natural Products Development and Innovative Drug Research,Beijing Union University,Beijing 100023,China)

机构地区：[1]北京联合大学功能因子与脑科学研究院 [2]北京联合大学天然产物开发与创新药物研究重点实验室,北京100023

出　　处：《天然产物研究与开发》2024年第11期1830-1837,共8页Natural Product Research and Development

基　　金：北京联合大学科研项目(ZKZD202304,ZK70202101)。

摘　　要：研究白藜芦醇(resveratrol,Res)对内质网应激途径细胞凋亡和糖原合成酶激酶-3β(glycogen synthase kinase-3β,GSK-3β)/Tau蛋白磷酸化作用的影响。体外原代培养神经元细胞,采用衣霉素(tunicamycin,TM)建立内质网应激模型,Western blot法检测内质网分子伴侣蛋白葡萄糖调节蛋白78(glucose-regulated protein 78,GRP78)、未折叠蛋白反应相关的肌醇需要酶1α(inositol-requiring enzyme 1α,IRE1α)的Ser724磷酸化、剪接的X盒结合蛋白1(spliced form of X-box binding protein 1s,XBP1s)表达、GSK-3β的Ser9和Tau蛋白的Ser396磷酸化水平。生物化学方法分析细胞质中半胱天冬酶-12(Caspase-12)和半胱天冬酶-3(Caspase-3)活性、细胞凋亡水平。结果显示,TM能够诱导内质网应激作用,导致神经元细胞GSK-3β的活化和Tau蛋白磷酸化水平升高(P<0.01)、神经元细胞经内质网途径凋亡(P<0.05)。与内质网应激抑制剂4-苯基丁酸结果相似,Res组显著降低了GRP78的表达(P<0.01)、降低了IRE1α-XBP1通路的活性(P<0.01)。Res可以减缓TM诱导的内质网途径细胞凋亡级联反应中Caspase-12和Caspase-3的活性(P<0.01)。Res抑制了TM诱导的GSK-3β的Ser9位点磷酸化水平和Tau蛋白Ser396位点的磷酸化水平(P<0.01)。结果表明,Res能够降低TM诱导的IRE1α-XBP1途径内质网应激作用、GSK-3β的活性及Tau蛋白发生磷酸化水平,减弱神经元细胞经内质网途径凋亡的级联反应作用。This study aims to investigate the effects of resveratrol(Res)on cell apoptosis through endoplasmic reticulum stress(ERS)pathway and phosphorylation of glycogen synthase kinase-3β(GSK-3β)and Tau protein.Primary cultured neurons were used in vitro to establish an ERS model induced by tunicamycin(TM).Western blot analysis was performed to detect the levels of the endoplasmic reticulum molecular chaperone protein glucose-regulated protein 78(GRP78),unfolded protein response related inositol-requiring enzyme 1α(IRE1α)phosphorylation and spliced form of X-box binding protein 1s(XBP1s)expression,phosphorylation of GSK-3βand Tau protein.Biochemical methods were used to analyze the activity of Caspase-12 and Caspase-3,as well as the level of cell apoptosis.The results indicated that TM induces endoplasmic reticulum stress,leading to increasing in neuronal GSK-3βactivation and Tau protein phosphorylation(P<0.01),as well as neuronal apoptosis through the endoplasmic reticulum pathway(P<0.05).Compared with the TM model group,like the inhibitor of endoplasmic reticulum stress 4-phenylbutyric acid,the Res protection group significantly reduced the expression of GRP78,phosphorylated IRE1αat Ser724,and XBP1s(P<0.01).Res reduced TM-induced the activity of both Caspase-12 and Caspase-3 and neuronal apoptosis(P<0.01).Res also inhibited TM-induced the phosphorylation levels of GSK-3βat Ser9 and Tau protein at Ser396(P<0.01).In conclusion,Res could reduce TM-induced endoplasmic reticulum stress on IRE1α-XBP1 pathway,the activity of GSK-3β,and the phosphorylation level of Tau protein,and weaken the cascade reaction of neuronal apoptosis through the endoplasmic reticulum pathway.

关 键 词：白藜芦醇 内质网应激 糖原合成酶激酶-3Β TAU蛋白 

分 类 号：R963[医药卫生—微生物与生化药学]