Correlation Research Of The Expression Of C-fos,Bcl-2,Caspase-3 Protein After Rat Neuron Mechanical Injuried With Apoptosis

Objective To study the expression of C-fos, Bcl-2, Caspase-3 protein and the relationship with neuron apoptosis after mechanical injury. Analyze the pathology and pathophysiology after acute brain trauma for improving the therapy.Method Take the cortical neuron from newborn rat and cultured. The cell was identified with microscope and munohistochemistry after 7 days in vitro cultivation maturely. The cell was injured mechanically by a10ul micropipet, light , moderate, sever group was created according to the number of strike. After that, cellular morphological changes were observed. In addition, cellular survival rate and lactate dehydrogenase (LDH) concentration were detected. Check the change of C-fos, Bcl-2, Caspase-3 protein in each group after injury by immunohistochemistry,and the cellular relative apoptosis ratio was detected by FCM(flow cytometre),and statistically analyzed the relationship between them.Results The percent of neurons was more than 90% according to the morphous and the immunohistochemistry at the 7 day. Apoptosis were discovered after mechanical injury. The cellular viability decreased and the LDH concentration increased after mechanical injury of neurons in vitro. C-fos, Bcl-2, Caspase-3 protein expressed in each injured group. There was significant difference between each group. The expression of C-fos, Bcl-2, Caspase-3 protein in severe group was larger than the one in light, moderate groups(P<0.05). C-fos protein expressed in one hour and reach peak at 24 hour.Bcl-2, Caspase-3 had a long duration time and arrived its peak at 48-72 hour. FCM showed the different apoptosis ratio after cell injury in each group. The apoptosis ratio in severe group was higher significantly than the one in light, moderate group(P<0.05).Conclusion In our study, cortical neurons were successfully cultured. It was confirmed that tip-caused injury could actually produce neuronal insults, and the degree of damage depended on the extension of tip-caused injury. C-fos, Caspase-3, Bcl-2 play a important role in the neuronal injury. They all participate in the neuronal apoptosis...