The Effect Of N6on The Cell Apoptosis After Spinal Cord Injury

PartⅠTHE EFFECT OF N6ON THE CELL APOPTOSIS AFTERSPINAL CORD INJURYObjective:To study the effect of N6(promote nerve regeneration factors)on apoptosis of nervecells after acute spinal cord injury.Methods:We performed the acute spinal cord injury with the NYU impactor under the Allen’smethod.The experimental animals were diveded into four groups,namely sham(A,onlyunder went laminectomy without weight-drop injury);SCI grou(B);SCI with NS treatedgroup(C);SCI with N6-treated group(D).The spinal cord tissues of the lesion epicenterwere dissected72hours after injury.Hematoxylin and eosin(HE)staining was used to showthe histopathological changes,and Caspase3was labeled through immuneohistochemistymethod;The expression of apoptosis-related gene Caspase3、Fas、Bax、Bcl2、P53wasmeasured by RT-PCR(reverse transcription polymerase chain reaction) and the expressionof apoptosis-related Bcl2with Western-Bloting.Results:The result of immunohistochemisty showed that Caspase3was expressed few in thenerve cells of A,but in the group of B and C it was expressed more than A(P<0.05) and Dexpressed Caspase3obviously fewer campared with B or C(P<0.05). From the result ofRT-PCR,we could found that the mRNA expression of caspase-3、Fas、P53is lowercampared group D with group B or C(P<0.05) while the mRNA expression of Bcl2/Baxwas higher in group D compared with group B or C(P<0.05) and the level of protein Bcl2like with the Bcl2/Bax. Conclusion:The promote nerve regeneration factors compound(N6) could inhibit the apoptosis ofnerve cells obviously in acute spinal cord injury. PartⅡSTUDY OF N6IN INHIBITING THE APOPTOSIS OFCULTURED SPINAL CORD NEURONS INDUCED BYGLUTAMIC ACIDObjective:To investigate the effect of promote nerve regeneration factors N6in inhibiting theapoptosis of cultured spinal cord neurons induced by glutamic acid.Methods:We cultured primary spinal cord neurons from13day old fetal rats and made theglutimic acid excitotoxicity model, N6was then added to protect neurons and the changesof cells were then investigated. AO/EB、Hochest and Tunel were used to detect the numberof the apoptosis of nerons,and CCK8was used to detect neurons viability. The study cellswere divided into four groups: the control group (C);Glu-injury group (Glu);Glu+NS treat-ed(NS);Glu+N6treated(N6).Cells of every guoup were cultivated in four dishes andrepeated three times.Result:Few apoptosis of neurons in group C,the apoptosis of neurons increased and theviability of neurons decreased obviously in group Glu and Ns.However,in group N6,theapoptosis of neurons decreased and the viability of neurons increased apparently comparedwith group Glu and NS(P<0.05).Conclution:Promote nerve regeration factors N6could inhibit the apoptosis of cultured spinalcord neurons inducted by glutamic acid.