Study On The Mechanism Of β-synuclein In Regulating Neuronal Injury And Repair

Objective This study clarified the expression of β-synuclein in spinal cord injury,and discussed the role and mechanism of β-synuclein in spinal cord neuron oxidative stress injury.Materials and methodsconstruct a spinal cord injury model of SD rat.The spinal cord tissues of the sham operation group and the spinal cord injury group were analyzed by protein mass spectrometry,immunohistochemistry,and q PCR to clarify the expression of β-synuclein in spinal cord injury.The rat β-synuclein gene was extracted by RT-PCR,and the eukaryotic recombinant plasmid GFP-β-synuclein was constructed by enzyme ligation and transformation.The interference fragment siβ-synuclein was constructed by si RNA,and GFP-β-synuclein and interference fragments were overexpressed in 293 T cells.The expression and interference efficiency of β-synuclein were verified by Western Blot.The primary hippocampal neurons of rats were cultured,and the GFP-β-synuclein plasmid and interference fragments were overexpressed with calcium phosphate transfection reagent.Image-Pro Plus was used to measure the length and number of branch points of hippocampal neurons.Construct SH-SY5 Y cell overexpressing β-synuclein,and add sodium L-glutamate to induce oxidative stress damage in SH-SY5 Y cells.CCK-8 reagent was used to detect the effect of β-synuclein on the oxidative stress damage of SH-SY5 Y induced by sodium L-glutamate.Results1)Using the Louisville Injury System Apparatus,SD rat spinal cord injury model was established;2)The results of protein mass spectrometry and principal component analysis on the spinal cord tissues of the sham operation group and the spinal cord injury group showed that β-synuclein is one of the most characteristic and most changing proteins in spinal cord injury;3)Immunohistochemistry and q PCR results showed that the expression of β-synuclein was down-regulated in SD rat with spinal cord injury.The more severe the spinal cord injury,the more obvious the down-regulation;4)The eukaryotic recombinant plasmid GFP-β-synuclein and the interference fragment siβ-synuclein were successfully constructed;5)Cultivate rat primary hippocampal neurons to overexpress GFP-β-synuclein plasmid and interference fragments.The results showed that overexpression of β-synuclein can promote the growth of rat hippocampal neurons,and interference with β-synuclein expression can inhibit the growth of rat hippocampal neurons;6)The oxidative stress damage model of SH-SY5 Y cells induced by glutamate was successfully established;7)The SH-SY5 Y cell overexpressing β-synuclein was successfully constructed,and sodium L-glutamate was added to induce the oxidative stress damage of SH-SY5 Y cells.Use CCK-8 reagent to detect cell viability.The results show that overexpression ofβ-synuclein can increase the cell survival rate of SH-SY5 Y cells induced by sodium L-glutamate,reduce the oxidative stress damage of SH-SY5 Y cells induced by sodium L-glutamate.Conclusions1)The expression of β-synuclein was down-regulated in SD rat with spinal cord injury.The more severe the spinal cord injury,the more obvious the down-regulation.2)β-synuclein can promote the growth of rat hippocampal neurons.3)β-synuclein can reduce cell oxidative stress damage induced by sodium L-glutamate.