Effect Of ?2-spectrin Protein Expression Level On Extent Of Damage To Spinal Cord Tissue

Purpose:To determine whether the level of 5hmC after SCI changes with the time of injury by detecting the overall 5hmC level in the peripheral blood of mice with different injury time.Detection of spinal cord tissue in the injured group and spinal cord tissue of the control group,and detection of changes in expression of the ?2-spectrin after inhibiting by Western blotDetection of changes in the extent of damage to spinal cord tissue by inhibiting the expression of ?2-spectrin protein Method:Establishment of a spinal cord injury model.28 ICR mice aged 5-6 weeks were selected and divided into 6 groups according to time points(1h,6h,12 h,24h,3d,7d).4 surgical mice and 4 sham-operated mice.The mice were sacrificed at the selected time points and the peripheral blood at the injury site was removed and stored at-80 °C.Peripheral blood DNA was extracted for Dot blot experiments.12 ICR mice aged 5-6 weeks were randomly divided into two groups,SCI group,SCI+7-NI group.1.SCI+7-NI group: Spinal cord tissue was exposed,and 0.1ml of ?2-spectrin protein inhibitor 7-NI(concentration: 1mg/ml)was Intraperitoneal injection administered with a 1ml syringe 5min prior to spinal cord injury,and spinal cord injury was modeled.2.7-NI group: After the spinal cord was exposed,only the drug was injected and the spinal cord tissue was not damaged.Western blot analysis was performed on the proteins of the spinal cord tissue from the SCI group and the SCI+7-NI group.The same method was used to establish four groups of models,SCI group,SCI+7-NI group,7-NI group,and control group.The above rat model was sacrificed at 24 h,the spinal cord tissue at the lesion was removed,and the-20 ° C refrigerator was quickly frozen and sectioned.Gross sections were observed using TTC staining.Result:Successfully produced a mouse model of spinal cord injury.The results showed that the 5hmC level of whole genome DNA was related to the injury time,reached the highest value at 24 h after injury.The results showed that protein expression of ?2-spectrin decreased after using 7-NI compared with the SCI group.The necrotic area after 7-NI treatment was larger than the necrotic area of other group.Conclusion:This study successfully established the SCI model of mice,and found that the level of 5hmC modification was significantly increased after spinal cord injury,peaked at 24 hours.Found 7-NI is the Inhibition of ?2-spectrin protein expression.Inhibition of ?2-spectrin protein expression aggravates spinal cord injury and neuronal cell death.