The Changes Of Proteome In The Spinal Cord Subjected To Partial Dorsal Root Rhizotmy And Acupuncture In Rats

[Objective] This study investigate the changs of proteome in the spinal cord of rats subjected to partial dorsal root rhizotomy and acupuncture .[Methods] The SD adult rats were divided into three groups randomly.Each Group consisted of 6 rats.The injured rats were subjected to surgical removal of the first lumbar through the fourth lumbar (L1-L4) and the sixth lumbar DRG,leaving the left L5 DRG untouched. The acupunctured rats received similar surgical treatment as Group injured,but in addition were subjected to acupuncture on the operated on the operated side at acupuncture points(xuewei穴位)zusanli-xuanzhong (足三里-悬钟) ,and futu-sanyinjiao (伏兔-三阴交) 7 days,once every day.The proteins in each group were respectively extracted from the lumber segment of spinal cord under anesthesia at 7 days post operation (dpo).Then the density of proteins were determined.Two dimensional polyacrylamide gel electrophoresis (2-DE) techniques were used to separate proteins;PDquest software was used to analyze their change of express in spinal cord.The mass-spectrum was applied to detected the peptide finger print map of differential expressed proteins .The bioinformatics were utilized to determined these differential expressed proteins. [Result] The results showed some reproducibility of proteins in 2-DE electrophoregram.The change of expression of the 12 kinds of proteins were preliminary screened and identified by PDquest soft and mass spectroscopy. The 12 kinds of proteins named form 1 to 12,known as Calmodulin , S-phase kinase-associated protein 1 A, synuclein,beta, Beta-synuclein, Cytochrome c oxidase polypeptide Va,mitochondrial precursor, peroxiredoxin 2, phosphatidylethanolamine binding protein, superoxide dismutase 1, Phosphoglycerate mutase 1, cofilin 1, peptidylprolyl isomerase A, Adenosine A2a receptor.The changes of expression of 12 proteins were significant among the sham operated and injuried and acupunctured in spinal cord in adult rats.The 3rd and 4th proteins were the same protein--Beta-synuclein. Another 4 kinds of proteins was special proteins, only showed up in some particular group.S-phase kinase-associated protein 1A only appeared in injuried group;cofilin 1 and Adenosine A2a receptor was not observed in the sham operated,merely wased observed in injuried and acupunctured,and the expression of cofilin 1 in acupunctured group descended instead of Adenosine A2a receptor rise.Peptidylprolyl isomerase A was not observed in the injuried,the expression of peptidylprolyl isomerase A up-regulation in acupunctured compare to in the sham operated.In addition, the expression of another 6 kinds of proteins and Beta-synuclein was down-regulated in injuried compare to in the sham operated, significant up-regulated in acupunctured contrast to injuried after operated.The 6 kinds of proteins are known as Calmodulin, Cytochrome c oxidase polypeptide Va, mitochondrial precursor,peroxiredoxin2, superoxide dismutase 1, phosphatidylethanolamine binding protein, Phosphoglycerate mutase 1．[Conclusion] Among the three groups,the reproducibility of electrophoregram were detected;the significant difference of express in Part of proteins were identified.This result suggests that these difference expressing proteins may participate in reparative process after spinal cord injury. The proteome technique mades up of 2-DE and mass-spectrum technique were established to detecte the change of expression of proteins in different tervention study.