中国危重病急救医学
中國危重病急救醫學
중국위중병급구의학
CHINESE CRITICAL CARE MEDICINE
2010年
8期
469-472,前插1
,共5页
高氧肺损伤%p38丝裂素活化蛋白激酶%N-乙酰半胱氨酸
高氧肺損傷%p38絲裂素活化蛋白激酶%N-乙酰半胱氨痠
고양폐손상%p38사렬소활화단백격매%N-을선반광안산
Hyperoxic lung injury%p38 mitogen-activated protein kinase%N-acetylcysteine
目的 观察p38丝裂素活化蛋白激酶(p38MAPK)信号途径在高氧肺损伤中的表达,探讨N-乙酰半胱氨酸(NAC)在高氧肺损伤中的保护作用及机制.方法 将30只幼年Wistar大鼠按随机数字表法分为空气对照组(A组)、高氧暴露组(B组)、高氧+NAC干预组(C组)、高氧+p38MAPK特异性抑制剂(SB203580)干预组(D组)、高氧+NAC+SB203580联合干预组(E组),每组6只.实验7 d后,光镜下观察肺组织病理改变,并行肺损伤评分;测定肺湿/干重(W/D)比值、支气管肺泡灌洗液(BALF)中总蛋白(TP)含量、肺通透系数;采用免疫组化法检测磷酸化p38MAPK(p-p38MAPK)在肺组织中的分布;用蛋白质免疫印迹法检测p-p38MAPK蛋白含量.结果 与A组比较,高氧各组均有不同程度的肺损伤,但药物干预各组(C、D、E组)肺损伤均较B组有所减轻.免疫组化显示,高氧各组p-p38MAPK阳性表达较A组明显增强,尤高表达在炎性浸润细胞;药物干预后(C、D、E组)p-p38MAPK阳性细胞较B组明显减少.蛋白质免疫印迹法显示,B组p-p38MAPK蛋白含量明显高于A组(0.20±0.03比0.11±0.01,P<0.05);干预后C、D、E组p-p38MAPK蛋白含量低于B组(0.16±0.02、0.15±0.01、0.14±0.02比0.20±0.03,均P<0.05),但仍高于A组(均P<0.05),而C、D、E组间则无明显差异.各组肺W/D比值、BALF中TP含量、肺通透系数改变与p-p38MAPK蛋白含量变化趋势一致.结论 高氧应激可激活损伤肺组织p38MAPK活性;NAC抗氧化肺保护作用机制可能是通过下调高氧诱导p38MAPK的激活而对肺损伤起保护作用.
目的 觀察p38絲裂素活化蛋白激酶(p38MAPK)信號途徑在高氧肺損傷中的錶達,探討N-乙酰半胱氨痠(NAC)在高氧肺損傷中的保護作用及機製.方法 將30隻幼年Wistar大鼠按隨機數字錶法分為空氣對照組(A組)、高氧暴露組(B組)、高氧+NAC榦預組(C組)、高氧+p38MAPK特異性抑製劑(SB203580)榦預組(D組)、高氧+NAC+SB203580聯閤榦預組(E組),每組6隻.實驗7 d後,光鏡下觀察肺組織病理改變,併行肺損傷評分;測定肺濕/榦重(W/D)比值、支氣管肺泡灌洗液(BALF)中總蛋白(TP)含量、肺通透繫數;採用免疫組化法檢測燐痠化p38MAPK(p-p38MAPK)在肺組織中的分佈;用蛋白質免疫印跡法檢測p-p38MAPK蛋白含量.結果 與A組比較,高氧各組均有不同程度的肺損傷,但藥物榦預各組(C、D、E組)肺損傷均較B組有所減輕.免疫組化顯示,高氧各組p-p38MAPK暘性錶達較A組明顯增彊,尤高錶達在炎性浸潤細胞;藥物榦預後(C、D、E組)p-p38MAPK暘性細胞較B組明顯減少.蛋白質免疫印跡法顯示,B組p-p38MAPK蛋白含量明顯高于A組(0.20±0.03比0.11±0.01,P<0.05);榦預後C、D、E組p-p38MAPK蛋白含量低于B組(0.16±0.02、0.15±0.01、0.14±0.02比0.20±0.03,均P<0.05),但仍高于A組(均P<0.05),而C、D、E組間則無明顯差異.各組肺W/D比值、BALF中TP含量、肺通透繫數改變與p-p38MAPK蛋白含量變化趨勢一緻.結論 高氧應激可激活損傷肺組織p38MAPK活性;NAC抗氧化肺保護作用機製可能是通過下調高氧誘導p38MAPK的激活而對肺損傷起保護作用.
목적 관찰p38사렬소활화단백격매(p38MAPK)신호도경재고양폐손상중적표체,탐토N-을선반광안산(NAC)재고양폐손상중적보호작용급궤제.방법 장30지유년Wistar대서안수궤수자표법분위공기대조조(A조)、고양폭로조(B조)、고양+NAC간예조(C조)、고양+p38MAPK특이성억제제(SB203580)간예조(D조)、고양+NAC+SB203580연합간예조(E조),매조6지.실험7 d후,광경하관찰폐조직병리개변,병행폐손상평분;측정폐습/간중(W/D)비치、지기관폐포관세액(BALF)중총단백(TP)함량、폐통투계수;채용면역조화법검측린산화p38MAPK(p-p38MAPK)재폐조직중적분포;용단백질면역인적법검측p-p38MAPK단백함량.결과 여A조비교,고양각조균유불동정도적폐손상,단약물간예각조(C、D、E조)폐손상균교B조유소감경.면역조화현시,고양각조p-p38MAPK양성표체교A조명현증강,우고표체재염성침윤세포;약물간예후(C、D、E조)p-p38MAPK양성세포교B조명현감소.단백질면역인적법현시,B조p-p38MAPK단백함량명현고우A조(0.20±0.03비0.11±0.01,P<0.05);간예후C、D、E조p-p38MAPK단백함량저우B조(0.16±0.02、0.15±0.01、0.14±0.02비0.20±0.03,균P<0.05),단잉고우A조(균P<0.05),이C、D、E조간칙무명현차이.각조폐W/D비치、BALF중TP함량、폐통투계수개변여p-p38MAPK단백함량변화추세일치.결론 고양응격가격활손상폐조직p38MAPK활성;NAC항양화폐보호작용궤제가능시통과하조고양유도p38MAPK적격활이대폐손상기보호작용.
Objective To investigate the expression of p38 mitogen-activated protein kinase (MAPK) in hyperoxic lung injury (HLI), and explore the protective effect of N-acetylcysteine (NAC) on HLI and its mechanism. Methods Thirty Wistar rats aged 3 weeks old were divided into five groups with 6 rats in each group according to random digits table: room-air group (A), hyperoxia injury group (B), hyperoxia+NAC group (C), hyperoxia+p38MAPK inhibitor (SB203580) group (D), hyperoxia+NAC+SB203580 group (E). Rats in NAC groups were injected with NAC (200 mg/kg) intraperitoneally, and they received an intravenous injection of SB203580 (0.5 mg/kg) in SB203580 groups. The animals were sacrificed after 7 days of experiment. Lung pathology and grade of lung tissue injury were examined with light microscopy, lung wet/dry (W/D) ratio, total protein (TP) level in bronchoalveolar lavage fluid (BALF) and permeability coefficient were evaluated. The location and quantity of phosphorylation p38MAPK (p-p38MAPK) protein were detected by immunohistochemistry and Western blotting analysis respectively. Results The pathological changes in the lung in B group included severe alveolar oedema with inflammatory cells aggregation and red blood cell leakage, while the lung pathological pictures in C, D, E groups were improved significantly compared with B group. p-p38MAPK positive cells increased in B group compared with those in A group, involving many types of pulmonary cells, especially in infiltrating inflammatory cells. In C, D, E groups, the positive cells remarkably decreased compared with B group. p-p38MAPK content was higher in B group than that in A group (0.20±0.03 vs. 0.11±0.01, P<0.05), and p-p38MAPK expressions in C, D, E groups decreased significantly compared with B group (0.16±0.02, 0.15±0.01, 0.14±0.02 vs. 0.20±0.03, all P<0.05), but were higher than those in A group (all P<0.05). There was no significant difference in p-p38MAPK quantity among three groups. Changes in W/D ratio, TP and permeability coefficient among groups were comparable with those of p-p38MAPK protein quantity. Conclusion Reactive oxygen species (ROS) activated p38MAPK signaling pathway. NAC may exert a protective effect on HLI through attenuation of hyperoxia-induced p38MAPK activation.
