Objective:To explore the mechanism of crocin,a major active component of Crocus sativus(Zanghonghua),in regulating amyloid beta(Aβ)generation,endoplasmic reticulum(ER)stress,and autophagy in neuronal cells,with poten...Objective:To explore the mechanism of crocin,a major active component of Crocus sativus(Zanghonghua),in regulating amyloid beta(Aβ)generation,endoplasmic reticulum(ER)stress,and autophagy in neuronal cells,with potential therapeutic applications in Alzheimer's disease(AD).Methods:Mouse neuroblastoma Neuron2a(N2a)cells stably transfected with the human amyloid precursor protein(APP)Swedish mutant was used as a cellular model for AD(N2a/APP).Control cells were vector transfected(N2a/vector).The effects of 3 different doses of crocin on reactive oxygen species(ROS)generation,cytosolic calcium,and apoptosis were evaluated by flow cytometry.Aβlevels were determined by enzyme-linked immunosorbent assay.APP processing and ER stress proteins expressions were determined by Western blot.Autophagosome formation was evaluated by autophagy detection kit and confocal microscope.Results:Crocin inhibited APP expression in N2a/APP cells and promotedα-cleavage of APP processing,while modestly reduced beta-secretase 1(BACE1)and presenilin 1(PS1,P<0.05 or P<0.01).ER stress markers,including the binding immunoglobulin protein/78-kD glucose-regulated protein(Bip/GRP78)and C/EBP homologous protein(CHOP),were elevated in N2a/APP cells compared to N2a/vector cells(P<0.05).Crocin could effectively reduce the levels of ER stress(P<0.05 or P<0.01).In addition,crocin enhanced autophagy by promoting formation of autophagosome(P<0.05 or P<0.01).Conclusion:Crocin significantly inhibited Aβgeneration by promotingα-cleavage of APP processing,inhibiting ER stressassociated unfolded protein response,and regulating autophagy.展开更多
基金Supported by the University of Macao (No.MYRG-2022-00248-ICMS).
文摘Objective:To explore the mechanism of crocin,a major active component of Crocus sativus(Zanghonghua),in regulating amyloid beta(Aβ)generation,endoplasmic reticulum(ER)stress,and autophagy in neuronal cells,with potential therapeutic applications in Alzheimer's disease(AD).Methods:Mouse neuroblastoma Neuron2a(N2a)cells stably transfected with the human amyloid precursor protein(APP)Swedish mutant was used as a cellular model for AD(N2a/APP).Control cells were vector transfected(N2a/vector).The effects of 3 different doses of crocin on reactive oxygen species(ROS)generation,cytosolic calcium,and apoptosis were evaluated by flow cytometry.Aβlevels were determined by enzyme-linked immunosorbent assay.APP processing and ER stress proteins expressions were determined by Western blot.Autophagosome formation was evaluated by autophagy detection kit and confocal microscope.Results:Crocin inhibited APP expression in N2a/APP cells and promotedα-cleavage of APP processing,while modestly reduced beta-secretase 1(BACE1)and presenilin 1(PS1,P<0.05 or P<0.01).ER stress markers,including the binding immunoglobulin protein/78-kD glucose-regulated protein(Bip/GRP78)and C/EBP homologous protein(CHOP),were elevated in N2a/APP cells compared to N2a/vector cells(P<0.05).Crocin could effectively reduce the levels of ER stress(P<0.05 or P<0.01).In addition,crocin enhanced autophagy by promoting formation of autophagosome(P<0.05 or P<0.01).Conclusion:Crocin significantly inhibited Aβgeneration by promotingα-cleavage of APP processing,inhibiting ER stressassociated unfolded protein response,and regulating autophagy.
