Sodium Ferulate Antagonizes Apoptosis Of Hippocampal Neurons Induced By Aβ(1-42) Through Notch1/Hes Signaling Pathway

ObjectiveDamage and loss of hippocampal nerve cells are the main causes of learning and memory impairment in patients with Alzheimer’s disease（AD）.At present,the mechanism has not been elucidated.Our previous in vitro and in vivo experiments proved that the Chinese medicine Chuanxiong and Angelica sinensis are effective.The monomeric component ferulic acid can resist hippocampal neuronal apoptosis induced by Aβ.Notch1 is a major signaling pathway that regulates apoptosis in the brain.It has been shown that in primary cultured mouse cortical neuron AD models,neurons Increased apoptosis is accompanied by upregulation of Notch1 expression.Based on previous research,this study uses Aβ_1-42to induce hippocampal neuron apoptosis,and further explores whether the protective effect of sodium ferulate（SF）is related to its regulation of the Notch1/Hes signaling pathway,providing an experiment for anti-AD treatment of SF in accordance with.MethodsTake the newborn SD rat fetal rats within 24 h,soak the suckling rats in 75%alcohol for 1 min,sever the heads and place them in sterile DMEM culture medium,remove the bilateral hippocampus under a microscope under sterile conditions,and disperse the cell suspension.Inoculated in a petri dish,and used for experiments after 7 days of culture.Dissolve Aβ_1-42in sterile saline in advance and incubate in a 37℃incubator for 48 h.The mature hippocampal neural cells are divided into the following four groups:control group（0.1%DMSO）for 72 h;Aβ_1-42group(Aβ_1-42,50 nmol/L)for 72 h;SF+Aβ_1-42group:SF（200μmol/L）for 6 h before Aβ_1-42;SF+Aβ_1-42+Jagged1 group（Jagged1 is a Notch1/Hes signal pathway activator）,SF（200μmol/L）and Jagged 1（300nmol/L）were added for 6 hours before Aβ_1-42.MTT was used to detect cell viability,TUNEL staining and flow cytometry to detect apoptosis.The relative protein expression of Notch1,Hes,Bax and Bcl-2 were detected by Western blot in each group.ResultsHippocampal neurons undergo a traumatic change 72 h after the addition of Aβ_1-42,showing that compared with the control group,the activity of hippocampal neurons in the Aβ_1-42injury group was significantly reduced,from100%±0.0%of the normal control group to 46.72±3.41%（P<0.05）,the percentage of apoptotic cells increased significantly compared with the normal control group.The TUNEL staining results showed an increase from normal1.12±0.63 to 16.05±1.03（P<0.01）.Flow cytometry showed that from the normal0.96±0.62 increased to 19.21±1.71（P<0.01）.The results of Western blot showed that the addition of Aβ_1-42significantly increased the expression of Notch1,Hes and Bax proteins in cultured hippocampal neurons in vitro,and significantly reduced the expression of Bcl-2 protein（P<0.05）.Compared with the Aβ_1-42group,the cell viability of the SF+Aβ_1-42group was significantly increased,the apoptosis rate was significantly reduced,the expressions of Notch1,Hes,and Bax proteins were significantly reduced,and the expression of Bcl-2 protein was significantly increased（P<0.05）.In order to explore the mechanism of SF,we applied Notch1/Hes signaling pathway agonist Jagged1,and found that after adding SF and Jagged1 for 6 hours,Aβ_1-42was added,which partially counteracted the neuronal apoptosis induced by SF on Aβ_1-42protection.ConclusionSF can inhibit the apoptosis of hippocampal neurons induced by Aβ_1-42,which may be related to the inhibition of Notch1/Hes signaling pathway.