CAJ | 학술논문

目的建立神经病理性痛大鼠脊髓蛋白质组图谱.方法成年雄性SD大鼠16只,体重260～320 g,随机分为2组(n=8):假手术组(S组)和神经病理性痛组(NP组).采用慢性缩窄性坐骨神经损伤法(CCI)制备大鼠神经病理性痛模型,S组仅暴露坐骨神经,不结扎.分别于CCI前1d(基础状态)及CCI后5、7 d测定机械痛阈和热痛阈;于CCI后7 d痛阈测定结束后取腰段脊髓组织,每组大鼠的脊髓组织混合后提取总蛋白质,测定脊髓组织蛋白质含量.以固相pH值梯度等电聚焦(IEF)为第一向,SDS-PAGE垂直电泳为第二向进行双向凝胶电泳(2-DE),制备脊髓蛋白质2-DE图谱,应用图像分析软件分析2-DE图谱,并观察其差异表达情况.结果与S组比较,NP组CCI后机械痛阈和热痛阈降低(P<0.05);两组大鼠脊髓组织总蛋白质经双向凝胶电泳分离,银染显色后得到分辨率高、重复性好的2-DE图谱,蛋白质点主要分布在pH 3～10,相对分子质量为8～120的区域;NP组差异表达的蛋白质有23个,其中16个蛋白质表达上调,7个蛋白质表达下调(P<0.05);表达上调幅度最大的为SSP2203蛋白,表达下调幅度最大的为SSP1807蛋白(P<0.05).结论本研究成功地建立了神经病理性痛大鼠脊髓蛋白质组的图谱.
목적 건립신경병이성통대서척수단백질조도보.방법 성년웅성SD대서16지,체중260～320 g,수궤분위2조(n=8):가수술조(S조)화신경병이성통조(NP조).채용만성축착성좌골신경손상법(CCI)제비대서신경병이성통모형,S조부폭로좌골신경,불결찰.분별우CCI전1d(기출상태)급CCI후5、7 d측정궤계통역화열통역;우CCI후7 d통역측정결속후취요단척수조직,매조대서적척수조직혼합후제취총단백질,측정척수조직단백질함량.이고상pH치제도등전취초(IEF)위제일향,SDS-PAGE수직전영위제이향진행쌍향응효전영(2-DE),제비척수단백질2-DE도보,응용도상분석연건분석2-DE도보,병관찰기차이표체정황.결과 여S조비교,NP조CCI후궤계통역화열통역강저(P<0.05);량조대서척수조직총단백질경쌍향응효전영분리,은염현색후득도분변솔고、중복성호적2-DE도보,단백질점주요분포재pH 3～10,상대분자질량위8～120적구역;NP조차이표체적단백질유23개,기중16개단백질표체상조,7개단백질표체하조(P<0.05);표체상조폭도최대적위SSP2203단백,표체하조폭도최대적위SSP1807단백(P<0.05).결론 본연구성공지건립료신경병이성통대서척수단백질조적도보.
Objective To establish the proteomies of the spinal cord in rats with neuropathie pain.Methods Sixteen adult male SD rats weighing 260-320 g Were randomly divided into 2 groups(n=8 each):sham operation group(S)and neuropathic pain group(NP).Neuropathie pain was induced by chronic constrictive injury(CCI).1he animals were anesthetized with intraperitoneal(IP)10%choral hydrate 300 mg/kg.The right sciatic nerve was exposed and 4 ligatures were placed on the sciafie nerve at 1 mm intervals.1heshold to thermal and mechanical stimulation was measured 1 day before and at 5,7 d after CCI.The animals were sacrificed at 7 d after pain threshold measurement.The lumbar enlargement segments of the spinal cord were harvested and proteins resolved by 2-DE.Differential proteome profiles were established and analyzed by means of immobilized pH gradient based on two-dimensional polyaerylamide gel electrophoresis(2D-PAGE).The 2-DE maps were visualized after silver staining and analyzed with image Master 2-DE software.Results There were 546 and 532 spots obtained in 2-DE maps of rat spinal cord tissue in group S and NP,and 23 proteins were found to be differentially expressed in NP group as compared with S group.Of the 23 proteins the expression of the 17 spots was increased and 6 spots decreased in quantity.Conclusion The optimized 2-DE profiles of proteome from the spinal cord of the 1"ats with neuropathie pain were established preliminarily.It lays,foundation for the proteome research of neuropathic pain.