部分背根切断大鼠脊髓蛋白质组学研究

发布时间：2018-08-19 17:34

【摘要】：目的：通过优化实验条件，建立双向电泳体系，初步获得正常大鼠脊髓蛋白质2—DE图谱。比较部分背根节切除14天后大鼠脊髓蛋白质与正常组间的表达差异，探讨脊髓可塑性的内在机制。方法：取成年SD雄性大鼠16只，，随机分为两组，每组8只。实验组8只SD大鼠切除双侧L_1—L_4和L_6背根节，保留L_5背根节；正常组8只SD大鼠不做任何特殊处理。模型动物于术后14天取材。采用双向电泳技术，初步分离蛋白质；采用质谱分析技术，比较部分背根节切除14天后大鼠脊髓蛋白质与正常组间蛋白质的肽指纹图和氨基酸序列组成差异。结果：建立了双向电泳体系，初步获得正常大鼠脊髓蛋白质在2—DE图谱从分子量大约10—97KD，等电点3—10之间800多个蛋白点。部分背根切断14天大鼠脊髓有3个与正常组比有差异表达的蛋白：(1)免疫球蛋白重链V区(Ig heavy chain V region(clone X41-2))，由127个aa组成，分子量(MW)为10.12KD，等电点(pI)为9.10。手术组该蛋白表达上调。(2)钙调蛋白A链复杂结构域(Chain A，CalmodulinIQ-Aa Domain Complex)，由148个aa组成，分子量为9.61KD，等电点为4.09。手术组该蛋白表达下调。(3)果糖二磷酸醛缩酶A(Fructose-bisphosphate aldolase A)，由364个aa组成的蛋白质，分子量36.36KD，等电点7.07，手术组该蛋白表达下调。结论：建立了双向电泳体系，初步获得正常大鼠脊髓蛋白质组学信息。发现部分去背根大鼠脊髓三个明显差异表达的蛋白质，三个差异表达的蛋白质可能与脊髓损伤及其可塑性机制有关。
[Abstract]:Aim: to establish a two-dimensional electrophoresis system to obtain the 2-DE map of normal rat spinal cord protein by optimizing the experimental conditions. To investigate the intrinsic mechanism of spinal cord plasticity by comparing the expression of spinal cord protein between rats and normal group 14 days after partial dorsal root ganglion resection. Methods: sixteen adult SD male rats were randomly divided into two groups, 8 rats in each group. The bilateral L_1-L_4 and L6 dorsal root ganglion were excised from 8 SD rats in the experimental group and L5 dorsal root ganglion was preserved, while 8 SD rats in the normal group were not given any special treatment. The model animals were selected 14 days after operation. Two-dimensional electrophoretic technique was used to isolate protein, and mass spectrometry was used to compare the peptide fingerprinting and amino acid sequence composition of protein between spinal cord protein and normal group of rats 14 days after partial dorsal root ganglion resection. Results: Two-dimensional electrophoretic system was established and more than 800 protein spots of normal rat spinal cord were obtained from molecular weight of 10-97 KD and isoelectric point 3-10 in 2-DE map. At 14 days after partial dorsal root dissection, there were three differentially expressed proteins in spinal cord of rats. (1) the immunoglobulin heavy chain V region (Ig heavy chain V region (clone X41-2), was composed of 127 AA. The molecular weight of (MW) is 10.12 KD and the isoelectric point (pI) is 9.10. (2) the complex domain of calmodulin A chain (Chain A Calmodulin IQ-Aa Domain Complex),) was composed of 148 AA, its molecular weight was 9.61 KD, and the isoelectric point was 4.09. (3) the protein of fructose diphosphate aldolase A (Fructose-bisphosphate aldolase A), with a molecular weight of 36.36 KD and isoelectric point of 7.07, was down-regulated in the operation group. Conclusion: a two-dimensional electrophoretic system was established and the proteomics information of normal rat spinal cord was preliminarily obtained. Three distinct differentially expressed proteins were found in the spinal cord of partially dorsal root removed rats, and the three differentially expressed proteins may be related to spinal cord injury and its plasticity.
【学位授予单位】：昆明医学院
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R341

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