大鼠横断性损伤坐骨神经后近端神经干分泌液的蛋白质组学分析和生物活性检测  

Proteomics analysis and biological detection of the secretion of proximal trunk of transected sciatic nerve in rats

作　　者：吕新生[1] 李新营[1] 肖志强[2] 李萃[2] 伍军[3] 

机构地区：[1]中南大学湘雅医院普外科,湖南长沙410008 [2]中南大学湘雅医院卫生部肿瘤蛋白质组学研究所,湖南长沙410008 [3]中南大学湘雅医院神经外科,湖南长沙410008

出　　处：《中国普通外科杂志》2004年第7期517-522,共6页China Journal of General Surgery

摘　　要：目的　探讨横断性周围神经近侧神经的分泌功能 ,分泌液的蛋白质成分及其对运动神经元生长的影响。方法　建立大鼠坐骨神经横断后神经近端室、神经远端室以及混合室 (近加远端神经 )收集室模型。用蛋白质双向电泳技术和质谱分析法分析 3种收集室内液体 (收集液 )的蛋白质成分 ,并以 3种收集液培养脊髓前角运动神经元。结果　大鼠坐骨神经断端的混合室、近端室和远端室均有淡黄色清亮液体 ,其蛋白质浓度分别为 ( 5 .45± 1.2 )mg/ml ,( 3 .87± 0 .7)mg/ml和 ( 5 .68± 1.5 )mg/ml。收集液蛋白质双向电泳结果显示 3组蛋白质图谱的图案模式相似。混合室、近端室和远端室组分别检测到 10 98± 3 4,985± 47和 10 2 1± 3 6个点。蛋白质点主要分布在pH4～ 8,分子量在2 5～ 90kD的范围内。应用 3组神经断端收集液培养运动神经元发现 ,3种收集液所培养的运动神经元的细胞存活数、神经元的胞体面积、突起长度总体上大于对照组 ;实验组之间无明显差异。结论　( 1)大鼠坐骨神经横断后 ,其近端神经干具有与远端神经干相似的分泌功能 ;( 2 )蛋白质双向电泳图谱的图案显示 3种收集室的蛋白质具有同源性 ;( 3 ) 3种收集液都能促进运动神经元的存活和生长 ,且近端室和远端室收集液的作用没有差别。Objective To study the protein components of the secretion of the proximal trunk of transected peripheral nerve and the effect of the secretion on the growth of motor neuron.Methods After the sciatic nerves of rats were transected, the proximal trunk secretion collecting compartment (P-SCC), distal trunk SCC (D-SCC) and (proximal) plus distal trunk SCC (PD-SCC) were set up. The protein components of the solution in the 3 types SCC were analyzed by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and mass spectrum. And the effect of 3 kinds solution on the growth of motor neuron was tested by the motor neuron culture respectively. (Results) All the P-SCC, D-SCC and PD-SCC were filled with light yellow, clear fluids. The protein (concentration) was (5.45±1.2)mg/ml,(3.87±0.7)mg/ml and (5.68±1.5)mg/ml in the solution of PD-SCC, P-SCC and D-SCC (respectively). The protein patterns of the 3 solution were similar, and the detected (protein) spots were 1098±34,985±47 and 1021±36 spots in the solution of P+D-SCC, P-SCC and D-SCC (respectively). Most of the protein spots (distributed) in the rangs of MV 25-90KD and PH 4-8. The results of motor neuron culture demonstrate that the number of survival cells, cell area, neurite length and OD value of experimental groups(including mix group, (proximal) group and distal group) are generally higher than control group. But there is no obvious difference between the three experimental groups. Conclusions (1)The proximal trunk of transected sciatic nerve had the similar secretive ability as the distal nerve trunk.(2)(Protein) image of the 3 kinds of SCC solution showed that proteins from the three SCC solution were homologous. (3)All the 3 kinds of SCC solution could promote the growth and activity of motor neuron, and there was no difference in function between P-SCC and D-SCC solution.

关 键 词：坐骨神经 分泌 蛋白质 分析 坐骨神经 损伤 

分 类 号：R322.85[医药卫生—人体解剖和组织胚胎学] Q51[医药卫生—基础医学]