Regulatory Efffct Of Macelignan On Hippocampal Neuron Damage Mediated By Glutamate

Objective:To explore the effect and regulation of Macelignan on glutamate-mediated hippocampal neuron HT22 cell damage.Methods:Using glutamate-induced hippocampal neuron damage cell model,by giving different concentrations of Macelignan to intervene to study its regulatory effect on neuronal damage and its mechanism.① MTT colorimetry was used to detect the effect of different concentrations of macelignan and glutamic acid on the activity of neuron.②The morphological changes of glutamate and myrissinol on the cells were observed by HE staining.③ The effects of glutamate and Macelignan on the expression of cyclinD1 and p21 were detected by immunohistochemistry.④The expression of cyclinD1 was detected by Western blot.⑤The apoptosis status was detected by Tunel method.⑥The expression of cyclins was detected by immunofluorescence staining.Result:1.The results of MTT assay showed that the rat hippocampal neuron HT22 cells were treated with the final concentration of 2.5,5,10,20 M Macelignan for 24 hours,and the survival rate was not significantly different from that of the control group,indicating that the final concentration of 2.5～20 M Macelignan had no toxic effect on the hippocampal neurons.Glutamate can reduce the survival rate of neurons,and with the intervention of Macelignan the survival rate of HT22 cells increased with the increase of Macelignan concentration.2.The results of HE staining showed that the hippocampal neurons in the control group grew polygonally and adhered roundly,with rounded nuclei and rounded cell bodies.The cells had multiple protrusions to form a synaptic network between cells.After 2.5mM glutamate treatment,the HT22 hippocampal neuron cell body was atrophy,and apoptotic bodies were found;the number of cells in the Macelignan composite treatment group was significantly increased compared with the glutamate treatment group,and the cells were more rounded and the proliferation was obvious,Indicating that Macelignan has obvious neuroprotective effect.3.The results of immunohistochemistry and western blot indicated that the expression of cyclinDl was significantly up-regulated in glutamate-mediated hippocampal neurons,while the expression was significantly inhibited after the intervention of Macelignan.4.Tunel method showed that a large number of fluorescent positive apoptotic cells could be found in the glutamate group,while the apoptotic cells were significantly reduced in the nutmeg intervention group,which was similar to the control group.5.Immunofluorescence results showed that Cleaved caspase3,CyclinDl,p21 and p27 were significantly increased in the glutamate group,while the number of positive cells in the glutamate and Maceligna treatment group was significantly reduced,which was similar to the control group and significantly inhibited the expression of cyclin and blocked the apoptosis of neurons.Conclusion:1.Macelignan can inhibit the neurotoxic effect mediated by glutamate and has obvious neuroprotective effect.2.Macelignan can inhibit the expression of p21,p27,cyclinD1,inhibit apoptosis and differentiation,and protect neurons.