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[严重烫伤后大鼠骨骼肌19S调节复合物的分离纯化及其蛋白水平变化]

[The isolation and purification of 19S regulator compound and the change of its protein level in rat skeletal muscle after severe scalding].

作者信息

Tan Yin-Ling, Wang Shi-Liang, Dong Yan-Lin

机构信息

Institute of Burn Research, Southwestern Hospital, The Third Military Medical University, Chongqing, 400038. P.R. China.

出版信息

Zhonghua Shao Shang Za Zhi. 2003 Feb;19(1):25-8.

Abstract

OBJECTIVE

To explore the role of 19S regulator compound protein in the degradation of skeletal muscle protein in scalded rats.

METHODS

Wistar rats were scalded and they were randomly divided into normal and 1, 2, 3, 5, and 7 postburn day (PBD) groups with 8 rats in each group. The 19S regulator compound of skeletal muscle in scalded rats was isolated and purified with chromatography. Rabbit-anti-rat antibody IgG of 19S regulator compound was prepared conventionally. The antibody was injected to rats in injection group (I) in which 19S antibody in dose of 3 mg/kg BW was injected for two times via tail vein with 6-hour interval. The rats in I group were decapitated on 1, 2 and 3 post-injection days, respectively. The scalded rats in control group (C) were treated in the same way, except that the antibody was replaced by normal saline. The change in content of 19S regulator compound was determined by western-blot. Meanwhile, the releasing rate of tyrosine from the skeletal muscle of scalded rats was also detected by fluorescent photography.

RESULTS

19S regulator compound with high purity was obtained. The content of 19S regulator compound in rat skeletal muscle was increased significantly after 2 PBD. But the protein degradation rate was also obviously increased on 2 PBD. The antibody of 19S compound might inhibit the enhancement of protein degradation.

CONCLUSION

Burn injury might up-regulate the protein level of skeletal muscle 19S regulator compound, which therefore activated the protein degradation by 26S protease compound. This might be an important factor leading to postburn negative nitrogen balance.

摘要

目的

探讨19S调节复合物蛋白在烫伤大鼠骨骼肌蛋白降解中的作用。

方法

将Wistar大鼠烫伤,随机分为正常组、烫伤后1、2、3、5和7天组,每组8只。采用层析法分离纯化烫伤大鼠骨骼肌的19S调节复合物。常规制备兔抗大鼠19S调节复合物抗体IgG。将该抗体注射到注射组(I组)大鼠体内,以3 mg/kg体重的剂量经尾静脉分两次注射19S抗体,间隔6小时。I组大鼠分别在注射后1、2和3天断头处死。对照组(C组)烫伤大鼠的处理方式相同,只是将抗体换成生理盐水。采用蛋白质免疫印迹法检测19S调节复合物含量的变化。同时,通过荧光摄影检测烫伤大鼠骨骼肌酪氨酸释放率。

结果

获得了高纯度的19S调节复合物。烫伤后2天,大鼠骨骼肌中19S调节复合物含量显著增加。但烫伤后2天蛋白质降解率也明显升高。19S复合物抗体可能抑制蛋白质降解的增强。

结论

烧伤可能上调骨骼肌19S调节复合物的蛋白水平,从而激活26S蛋白酶体复合物介导的蛋白质降解。这可能是导致烧伤后负氮平衡的重要因素。

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