【摘要】：目的：通過(guò)優(yōu)化實(shí)驗(yàn)條件，建立雙向電泳體系，初步獲得正常大鼠脊髓蛋白質(zhì)2—DE圖譜。比較部分背根節(jié)切除14天后大鼠脊髓蛋白質(zhì)與正常組間的表達(dá)差異，探討脊髓可塑性的內(nèi)在機(jī)制。 方法：取成年SD雄性大鼠16只，，隨機(jī)分為兩組，每組8只。實(shí)驗(yàn)組8只SD大鼠切除雙側(cè)L_1—L_4和L_6背根節(jié)，保留L_5背根節(jié)；正常組8只SD大鼠不做任何特殊處理。模型動(dòng)物于術(shù)后14天取材。采用雙向電泳技術(shù)，初步分離蛋白質(zhì)；采用質(zhì)譜分析技術(shù)，比較部分背根節(jié)切除14天后大鼠脊髓蛋白質(zhì)與正常組間蛋白質(zhì)的肽指紋圖和氨基酸序列組成差異。 結(jié)果：建立了雙向電泳體系，初步獲得正常大鼠脊髓蛋白質(zhì)在2—DE圖譜從分子量大約10—97KD，等電點(diǎn)3—10之間800多個(gè)蛋白點(diǎn)。部分背根切斷14天大鼠脊髓有3個(gè)與正常組比有差異表達(dá)的蛋白：(1)免疫球蛋白重鏈V區(qū)(Ig heavy chain V region(clone X41-2))，由127個(gè)aa組成，分子量(MW)為10.12KD，等電點(diǎn)(pI)為9.10。手術(shù)組該蛋白表達(dá)上調(diào)。(2)鈣調(diào)蛋白A鏈復(fù)雜結(jié)構(gòu)域(Chain A，CalmodulinIQ-Aa Domain Complex)，由148個(gè)aa組成，分子量為9.61KD，等電點(diǎn)為4.09。手術(shù)組該蛋白表達(dá)下調(diào)。(3)果糖二磷酸醛縮酶A(Fructose-bisphosphate aldolase A)，由364個(gè)aa組成的蛋白質(zhì)，分子量36.36KD，等電點(diǎn)7.07，手術(shù)組該蛋白表達(dá)下調(diào)。 結(jié)論：建立了雙向電泳體系，初步獲得正常大鼠脊髓蛋白質(zhì)組學(xué)信息。發(fā)現(xiàn)部分去背根大鼠脊髓三個(gè)明顯差異表達(dá)的蛋白質(zhì)，三個(gè)差異表達(dá)的蛋白質(zhì)可能與脊髓損傷及其可塑性機(jī)制有關(guān)。
[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.
【學(xué)位授予單位】：昆明醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2006
【分類號(hào)】：R341

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