大鼠骨骼肌26S蛋白酶复合体在严重烫伤后含量的变化

The changes of 26S proteasome level in skeletal muscle of the severely scalding rats

目的 为阐明及防治烧伤后负氮平衡提供理论依据。方法 层析技术结合盐析和离心分离、纯化 2 6S蛋白酶复合体 ,常规制备抗 2 6S蛋白酶复合体的抗体IgG ,用固相免疫印迹法测定 2 6S蛋白酶复合体含量的变化 ,用荧光分光光度法测定骨骼肌中酪氨酸的释放速率。结果 获得纯度较高的 2 6S蛋白酶复合体。从烫伤后第 2天起 ,大鼠骨骼肌中的 2 6S蛋白酶复合体便开始明显增加 ,而骨骼肌蛋白质降解速率也在伤后第 2天显著增强 ,但 2 6S抗体则能明显抑制蛋白质降解的增加。结论 烧伤刺激可上调骨骼肌 2 6S蛋白酶复合体的蛋白水平 ,从而增强蛋白质的降解作用 。

Objective To support the theoretical base of the negative nitrogen balance. Methods The chromatography coupled with fractionated salting out and centrifugation were applied to purify the 26S proteasome.The rabbit anti rat 26S proteasome IgG was obtained by conventional means.And the content of 26S proteasome was determined by western blot. Meanwhile,the release of tyrosine was assayed by fluorescentphotography.Results The comparetivly purified 26S proteasome was obtained .The contents of 26S protesome in rat skeletal muscle markedly incresed from the second day after scalded, The rate of protein degradation was also enchanced,which the 26S proteasome IgG could inhibit.Conclusion Burn can stimulate the protein level of 26S proteasome in skeletal muscle, thus enhance the degradation of protein, which is associated with the development of negative nitrogen balance following scald