中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2012年
12期
2527-2530
,共4页
耿庆%王贤灿%杨晓辉%刘军韬%胡浩%M.Majetschak
耿慶%王賢燦%楊曉輝%劉軍韜%鬍浩%M.Majetschak
경경%왕현찬%양효휘%류군도%호호%M.Majetschak
肺损伤%泛素%蛋白酶体
肺損傷%汎素%蛋白酶體
폐손상%범소%단백매체
Lung injure%Ubiquitin%Proteasome
目的 探讨支气管肺泡腔等细胞外液中是否存在泛素蛋白酶体及其在急性肺损伤过程中的变化和作用机制.方法 应用大鼠肺挫伤模型研究急性肺损伤支气管肺泡腔内泛素蛋白酶体含量变化及其可能的病理生理作用.分别采用生化检测、酶联免疫吸附试验(ELISA)及Western blot等方法研究对照组及急性肺损伤组伤后6～168 h支气管肺泡灌洗液(BALF)中蛋白酶体20S亚基、泛素等含量及其活性.结果 急性肺损伤后其BALF中20S蛋白酶体、泛素含量明显增加,在24 h达到峰值(20S:(2397 ±71) μg/L;泛素(469±124) μg/L),168 h恢复至正常水平(20S:(42±30) μg/L;泛素(31±6) μg/L).BALF中ATP/ Mg2+依赖的多肽酶Epoxomicin敏感的胰凝乳蛋白酶样(CT-L)及Epoxomicin敏感的胰蛋白酶样(T-L)酶活性在肺挫伤后6～24 h明显增加,两者均在伤后24 h增高到峰值[CT-L:(3621±245) pmol/(h·ml);T-L:(736±91) pmol/(h·ml)].在急性肺损伤的BALF中加入ATP/ Mg2+可导致蛋白质降解明显增加,但加入Epoxomicin或乙二胺四乙酸(EDTA)可显著抑制蛋白质降解.结论 蛋白酶体存在于细胞外液;在急性肺损伤后,蛋白酶体释放进入细胞外支气管肺泡腔中,在肺泡蛋白质降解、廓清过程中起重要作用.
目的 探討支氣管肺泡腔等細胞外液中是否存在汎素蛋白酶體及其在急性肺損傷過程中的變化和作用機製.方法 應用大鼠肺挫傷模型研究急性肺損傷支氣管肺泡腔內汎素蛋白酶體含量變化及其可能的病理生理作用.分彆採用生化檢測、酶聯免疫吸附試驗(ELISA)及Western blot等方法研究對照組及急性肺損傷組傷後6～168 h支氣管肺泡灌洗液(BALF)中蛋白酶體20S亞基、汎素等含量及其活性.結果 急性肺損傷後其BALF中20S蛋白酶體、汎素含量明顯增加,在24 h達到峰值(20S:(2397 ±71) μg/L;汎素(469±124) μg/L),168 h恢複至正常水平(20S:(42±30) μg/L;汎素(31±6) μg/L).BALF中ATP/ Mg2+依賴的多肽酶Epoxomicin敏感的胰凝乳蛋白酶樣(CT-L)及Epoxomicin敏感的胰蛋白酶樣(T-L)酶活性在肺挫傷後6～24 h明顯增加,兩者均在傷後24 h增高到峰值[CT-L:(3621±245) pmol/(h·ml);T-L:(736±91) pmol/(h·ml)].在急性肺損傷的BALF中加入ATP/ Mg2+可導緻蛋白質降解明顯增加,但加入Epoxomicin或乙二胺四乙痠(EDTA)可顯著抑製蛋白質降解.結論 蛋白酶體存在于細胞外液;在急性肺損傷後,蛋白酶體釋放進入細胞外支氣管肺泡腔中,在肺泡蛋白質降解、廓清過程中起重要作用.
목적 탐토지기관폐포강등세포외액중시부존재범소단백매체급기재급성폐손상과정중적변화화작용궤제.방법 응용대서폐좌상모형연구급성폐손상지기관폐포강내범소단백매체함량변화급기가능적병리생리작용.분별채용생화검측、매련면역흡부시험(ELISA)급Western blot등방법연구대조조급급성폐손상조상후6～168 h지기관폐포관세액(BALF)중단백매체20S아기、범소등함량급기활성.결과 급성폐손상후기BALF중20S단백매체、범소함량명현증가,재24 h체도봉치(20S:(2397 ±71) μg/L;범소(469±124) μg/L),168 h회복지정상수평(20S:(42±30) μg/L;범소(31±6) μg/L).BALF중ATP/ Mg2+의뢰적다태매Epoxomicin민감적이응유단백매양(CT-L)급Epoxomicin민감적이단백매양(T-L)매활성재폐좌상후6～24 h명현증가,량자균재상후24 h증고도봉치[CT-L:(3621±245) pmol/(h·ml);T-L:(736±91) pmol/(h·ml)].재급성폐손상적BALF중가입ATP/ Mg2+가도치단백질강해명현증가,단가입Epoxomicin혹을이알사을산(EDTA)가현저억제단백질강해.결론 단백매체존재우세포외액;재급성폐손상후,단백매체석방진입세포외지기관폐포강중,재폐포단백질강해、곽청과정중기중요작용.
Objective To explore the presence and role of ubiquitin-proteasome-pathway (UPP) in the bronchoalveolar space of the injured lung.Methods A rat model of lung contusion (LC) was used to evaluate the pathophysiological role of UPP in the injured lung.Fifty SD rats were divided into control groups and lung contusion group (LC).Bronchoalveolar lavage fluids (BALF) were obtained at different time points up from 6 h to 168 h after LC or control groups.The concentration of protein,enzyme activities,enzyme linked immunosorbent assay (ELISA) and Western blotting were used to evaluate the UPP in the BALF.Results The data indicated the presence of ubiquitin and proteasomes in BALF,20S and ubiquitin were increased significantly after LC,peaked at 24 h [20S:(2397 ±71) μg/L; ubiquitin (469 ±124) μg/L] and normalized at 168 h [20S:(42 ±30) μg/L; ubiquitin (31 ±6) μg/L) after LC.Mg2+/ATP-dependent peptidase activities were detected 6-24 h in BALF afer LC,and peaked at 24 h[CT-L:(3621 ±245) pmol x h-1 x mL-1; T-L:(736 ±91) pmol ×h-1 ×mL-1).Addition of Mg2+/ATP in BALF accelerated the proteolysis,which could be prevented by Epoxomicin and EDTA.Conclusion The UPP exists in the extracellular space.The proteasomes is released in BALF after LC,which may play an important role in injured lung.
