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。
Resveratrol inhibits tunicamycin-induced neuronal apoptosis and phosphorylation of GSK-3β/Tau protein through IRE1α-XBP1 pathway
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 neu-rons were used in vitro to establish an ERS model induced by tunicamycin(TM).Western blot analysis was performed to de-tect the levels of the endoplasmic reticulum molecular chaperone protein glucose-regulated protein 78(GRP78),unfolded pro-tein response related inositol-requiring enzyme 1α(IRE1α)phosphorylation and spliced form of X-box binding protein 1s(XBP1s)expression,phosphorylation of GSK-3β and 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-3β activation 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 IRE1α at 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-3β at 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 neu-ronal apoptosis through the endoplasmic reticulum pathway.
resveratrolendoplasmic reticulum stressglycogen synthase kinase-3βTau protein
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维普
