OBJECTIVE:To explore the protective mechanism of spinosin(SPI)on Alzheimer's disease(AD)model cells,Neuro-2a/APP695(N2a/APP695),against H_(2)O_(2)-induced oxidative stress damage,to reflect the influence of oxidat...OBJECTIVE:To explore the protective mechanism of spinosin(SPI)on Alzheimer's disease(AD)model cells,Neuro-2a/APP695(N2a/APP695),against H_(2)O_(2)-induced oxidative stress damage,to reflect the influence of oxidative stress on the development of AD,and to provide a valuable basis for the research and development of therapeutic drug for AD.METHODS:N2a/APP695 cells were exposed to H2O2 and then treated with spinosin.Firstly,the secretion level of amyloidβ(Aβ)1-42 and the production of malondialdehyde(MDA)and lactate dehydrogenase(LDH)were detected by enzyme linked immunosorbent assay kits.Secondly,the oligomerization degree of Aβ1-42 was performed by Thioflavin T staining.Thirdly,the expression levels of p-Tau(Ser199/202/396),synaptophysin(SYP),postsynaptic density protein 95(PSD95),and mitogen-activated protein kinase(MAPK)family-related proteins were detected by Western blot analysis.In addition,FITC-labeled phalloidin was used in cytoskeleton staining to reflect synaptic function.RESULTS:This study showed that H2O2 stimulated N2a/APP695 cells to produce excessive MDA and LDH and secrete a large amount of Aβ,promoted the aggregation of Aβ,induced Tau protein hyperphosphorylation,and led to synaptic dysfunction.Spinosin reversed these changes caused by H2O2 by inactivating p38,which was verified by treatment with the p38 inhibitor BIRB796.CONCLUSION:Spinosin protects N2a/APP695 cells from oxidative stress damage caused by H2O2 through inactivating p38.展开更多
基金National Natural Science Foundation of China:Study on the Basis and Mechanism of Antidepressant Substances in Which Schisandra Chinensis Regulates the Endocannabinoid System in the Body and Participates in"Gut-Brain Dialogue"(No.82173961)National Natural Science Foundation of China:to Investigate the Pharmacological Substance Basis and Mechanism of Suanzaoren Decoction in Improving Depression by Crosstalk Regulating"Gut Microbiota-Gut-Brain"Axis Based on TLR4-NF-κB/NLRP3 and Wnt/β-Catenin Signaling Pathways(No.82003926)+2 种基金the Doctoral Scientific Research Foundation of Liaoning Province:Study on the Mechanism of Antidepressant Action of Schisandra Chinensis Mediated by Intestinal Flora(2019-BS-233)High-Level Innovation and Entrepreneurship Team of Liaoning Province:"Healthy Liaoning"Functional Food Quality and Safety Innovation Team,"Xingliao Talent Plan"High-Level Innovation and Entrepreneurship Team(XLYC2008029)Guizhou Provincial Natural Science Foundation:Probing the Chemical Components and the Detailed Working Mechanisms of Danggui Buxue Tang for Mitigating the Menopausal Syndrome(QKH-J[2020]1Y377)。
文摘OBJECTIVE:To explore the protective mechanism of spinosin(SPI)on Alzheimer's disease(AD)model cells,Neuro-2a/APP695(N2a/APP695),against H_(2)O_(2)-induced oxidative stress damage,to reflect the influence of oxidative stress on the development of AD,and to provide a valuable basis for the research and development of therapeutic drug for AD.METHODS:N2a/APP695 cells were exposed to H2O2 and then treated with spinosin.Firstly,the secretion level of amyloidβ(Aβ)1-42 and the production of malondialdehyde(MDA)and lactate dehydrogenase(LDH)were detected by enzyme linked immunosorbent assay kits.Secondly,the oligomerization degree of Aβ1-42 was performed by Thioflavin T staining.Thirdly,the expression levels of p-Tau(Ser199/202/396),synaptophysin(SYP),postsynaptic density protein 95(PSD95),and mitogen-activated protein kinase(MAPK)family-related proteins were detected by Western blot analysis.In addition,FITC-labeled phalloidin was used in cytoskeleton staining to reflect synaptic function.RESULTS:This study showed that H2O2 stimulated N2a/APP695 cells to produce excessive MDA and LDH and secrete a large amount of Aβ,promoted the aggregation of Aβ,induced Tau protein hyperphosphorylation,and led to synaptic dysfunction.Spinosin reversed these changes caused by H2O2 by inactivating p38,which was verified by treatment with the p38 inhibitor BIRB796.CONCLUSION:Spinosin protects N2a/APP695 cells from oxidative stress damage caused by H2O2 through inactivating p38.
