Research About The Neuro-protective Effect During Different Time Courses On SCI Rats Treated By Jiaji Electro-Acupuncture Of Different Frequency

Objective:To determine the effects of electroacupuncture (EA) treatment with different waves on the apoptosis and related factors including Caspase-3 and Bcl-2?Bax expression at protein and gene levels; while and the damage repair related factors including NGF, BDNF, and VEGF expression of spinal cord injury (SCI) rats, and to explore the neuro-protective mechanism in programmed cell death of SCI rats treated with EA, which provides a new idea and basic theory for clinical treatment of SCI.Methods:One hundred and sixty-five healthy SD rats were randomly and equally divided into Sham group, Model group, Methylprednisolone group, Rarefaction Wave group and Rarefaction-Dense Wave group, each group 33 rats.Each single group was also randomly divided into 3 time units (8h,3d,7d),11 rats in each unit. SCI model was induced by modified Allen's method.The morphologic change, cell apoptosis and ultrastructure changes of injured spinal cord were observed by HE, Transmissive Electronic Microscope and TUNEL methods.The mRNA and protein expressions changes of Caspase-3 and Bcl-2, Bax in spinal cord nerve cells were observed by RT-PCR, Western Blot, the expression of NGF, BDNF, and VEGF in the spinal cord injury area were tested by immunohistochemical method.And.The experiment data were analyzed by Image Analysis systerm and the SPSS 17.0, Computer Linked Image Analysis and then obtained the results.Results:1.Morphological changes:The tissue damage of sham group are changes few, the three treatment groups are much more serious, while the model group is the most serious. The classical changes in the model and three intervention groups are as follows:neuron cell appears edema, neuron vacuolated and the infiltration of the inflammatory cells.2.Ultrastructural Change of neuron:Cell morphological changes are as follows:the cytoplasm Shrinkaged or contracted, nuclear membrane enveloped, heterochromatin margined and cell nucleus condensed; Cell, Mitochondria and Endoplasmic Reticulum (ER) all swelled and ER membrane dissolved to various degrees. The nerve damaged in the sham group was not significantly changed, while the model and other three treatment groups showed gradual seriousness with prolongation of duration; the moder group was worst, the other three treatment groups were much better.3.TUNELStaining:The classical changes of model and three treatment groups are as follows:cell chromatin condensed and aggregated spot-like along the nuclear membrane edge with cytoplasm hyperchromia. There are not significant changes in the sham group.4.Western Blot and RT-PCR:(1)After spinal cord injury, the mRNA and protein expressions of Caspase-3 and Bax during different time course in the model group increased while that of the three treatment groups decreased with the obvious difference comparing with the model group. Among the three treatment groups, the data of different time course of Rarefaction Wave group is the lowest, and that of Rarefaction-Dense Wave group is lower than Methylprednisolone group.(2)After spinal cord injury, the mRNA and protein expressions of Bcl-2 during different time course in the model group increased while that of the three treatment groups increased apparently with the obvious difference comparing with the model group. Among the three treatment groups, the data of different time course of Rarefaction Wave group is the highest, and that of Rarefaction-Dense Wave group is higher than Methylprednisolone group.5.Immunohistochemical method:After spinal cord injury, the expressions of NGF, BDNF, and VEGF during different time course in the model group increased while that of the three treatment groups increased apparently with the obvious difference comparing with the model group. Among the three treatment groups, the data of different time course of Rarefaction Wave group is the highest, and that of Rarefaction-Dense Wave group is higher than Methylprednisolone group.Conclusion:1.After spinal cord injury, both of the electro-acupuncture and Methylprednisolone confront the apoptosis by reducing the expression of apoptosis starts factor Caspase-3, elevating the expression of apoptosis suppression factor Bcl-2 while lowering that of apoptosis promotion factor Bax.2.After spinal cord injury, both of the electro-acupuncture and Methylprednisolone promote the spinal cord repair by increasing the expression of nerve growth factors including NGF, BDNF, and VEGF.3.Jiaji EA can inhibit cell apoptosis in some degrees after spinal cord injury, and has some neuro-protective effects. However, it can not inhibit cell apoptosis comprehensively.4.After spinal cord injury, Jiaji EA play the comprehensive neuro-protective effects of multiple targets and ways by the inhibition of apoptosis process on the one hand, and activation of the nerves and blood vessels reparation on the other hand with the effect of Rarefaction Wave better than Rarefaction-Dense Wave.