中华劳动卫生职业病杂志
中華勞動衛生職業病雜誌
중화노동위생직업병잡지
CHINESE JOURNAL OF INDUSTRIAL HYGIENE AND OCCUPATIONAL DISEASES
2012年
4期
278-283
,共6页
邵义如%申捷%袁震%何岱昆%张琳
邵義如%申捷%袁震%何岱昆%張琳
소의여%신첩%원진%하대곤%장림
光气%丝裂原活化蛋白激酶%肺损伤
光氣%絲裂原活化蛋白激酶%肺損傷
광기%사렬원활화단백격매%폐손상
Phosgene%Mitogen-activated protein kinase(MAPK)%Lung injury
目的 探讨丝裂原活化蛋白激酶(mitogen activated protein kinases,MAPKs)的亚通路信号蛋白-细胞外信号调节蛋白激酶(extracellular signal-regulated kinase,ERK1/2)、P38和c-氨基末端蛋白激酶(c-Jun N-terminal kinase,JNK)在大鼠光气吸入肺损伤组织中的表达及各通路在光气吸入性肺损伤中的作用.方法 选取30只雄性Wistar大鼠随机分成空气对照组、光气吸入组、PD98059(ERK1/2特异性阻断剂)、SB203580(P38特异性阻断剂)和SP600125(JNK特异性阻断剂)干预组,每组6只,应用定向流动光气吸入装置,复制大鼠光气(8.33 L/mg)吸入性肺损伤的模型,空气对照组吸入空气,3个干预组均在光气(8.33 L/mg)吸入前给予PD98059、SB203580、SP600125.分别采用免疫组化及蛋白质印迹( Western blot)法检测肺组织MAPKs(ERK1/2、P38、JNK)蛋白及磷酸化蛋白p-MAPKs (p-ERK 1/2、p-P38、p-JNK)的定性定量表达.观察肺组织病理学改变、计数支气管灌洗液(BALF)中中性粒细胞数、测定肺湿干比.结果 空气对照组仅肺泡上皮细胞及气道上皮细胞见少量p-ERK1/2、p-P38、p-JNK阳性表达；光气吸入组p-ERK1/2、p-P38、p-JNK阳性细胞数明显增多,广泛分布于肺内细胞:肺泡上皮细胞、气道上皮细胞、胸膜间皮细胞、浸润炎细胞及间质纤维细胞；干预组p-ERK1/2、p-P38、p-JNK阳性细胞数减少.各组ERK、P38、JNK的表达无明显变化；光气吸入组较空气对照组p-P38、p-JNK蛋白表达增强,3个干预组的p-ERK1/2、p-P38、p-JNK的蛋白表达水平均低于光气吸入组,差异均有统计学意义(P＜0.05,P＜0.01).与空气对照组[中性粒细胞计数:(2.05±0.75)×104,肺湿干比:(3.64％±0.72％)]比较,光气吸入组肺损伤程度严重,表现出肺损伤的典型病理学特征,且BALF中中性粒细胞数[(10.78±2.16)×104]增多,肺湿干比(7.65％±0.58％)升高,SP600125干预组和SB203580干预组BALF中中性粒细胞数[SP600125组:(7.86±2.12)×104,SB203580组:(8.43±1.51)×104]和肺湿干比[SP600125组:(6.10％±0.97％),SB203580组:(6.09％±1.43％)]降低,差异均有统计学意义(P＜0.05,P＜0.01).结论 光气吸入可能激活MAPK信号通路,通过p-MAPKs表达增加在光气吸入后的肺损伤中发挥作用,其中以p-P38、p-JNK活性变化为主.
目的 探討絲裂原活化蛋白激酶(mitogen activated protein kinases,MAPKs)的亞通路信號蛋白-細胞外信號調節蛋白激酶(extracellular signal-regulated kinase,ERK1/2)、P38和c-氨基末耑蛋白激酶(c-Jun N-terminal kinase,JNK)在大鼠光氣吸入肺損傷組織中的錶達及各通路在光氣吸入性肺損傷中的作用.方法 選取30隻雄性Wistar大鼠隨機分成空氣對照組、光氣吸入組、PD98059(ERK1/2特異性阻斷劑)、SB203580(P38特異性阻斷劑)和SP600125(JNK特異性阻斷劑)榦預組,每組6隻,應用定嚮流動光氣吸入裝置,複製大鼠光氣(8.33 L/mg)吸入性肺損傷的模型,空氣對照組吸入空氣,3箇榦預組均在光氣(8.33 L/mg)吸入前給予PD98059、SB203580、SP600125.分彆採用免疫組化及蛋白質印跡( Western blot)法檢測肺組織MAPKs(ERK1/2、P38、JNK)蛋白及燐痠化蛋白p-MAPKs (p-ERK 1/2、p-P38、p-JNK)的定性定量錶達.觀察肺組織病理學改變、計數支氣管灌洗液(BALF)中中性粒細胞數、測定肺濕榦比.結果 空氣對照組僅肺泡上皮細胞及氣道上皮細胞見少量p-ERK1/2、p-P38、p-JNK暘性錶達；光氣吸入組p-ERK1/2、p-P38、p-JNK暘性細胞數明顯增多,廣汎分佈于肺內細胞:肺泡上皮細胞、氣道上皮細胞、胸膜間皮細胞、浸潤炎細胞及間質纖維細胞；榦預組p-ERK1/2、p-P38、p-JNK暘性細胞數減少.各組ERK、P38、JNK的錶達無明顯變化；光氣吸入組較空氣對照組p-P38、p-JNK蛋白錶達增彊,3箇榦預組的p-ERK1/2、p-P38、p-JNK的蛋白錶達水平均低于光氣吸入組,差異均有統計學意義(P＜0.05,P＜0.01).與空氣對照組[中性粒細胞計數:(2.05±0.75)×104,肺濕榦比:(3.64％±0.72％)]比較,光氣吸入組肺損傷程度嚴重,錶現齣肺損傷的典型病理學特徵,且BALF中中性粒細胞數[(10.78±2.16)×104]增多,肺濕榦比(7.65％±0.58％)升高,SP600125榦預組和SB203580榦預組BALF中中性粒細胞數[SP600125組:(7.86±2.12)×104,SB203580組:(8.43±1.51)×104]和肺濕榦比[SP600125組:(6.10％±0.97％),SB203580組:(6.09％±1.43％)]降低,差異均有統計學意義(P＜0.05,P＜0.01).結論 光氣吸入可能激活MAPK信號通路,通過p-MAPKs錶達增加在光氣吸入後的肺損傷中髮揮作用,其中以p-P38、p-JNK活性變化為主.
목적 탐토사렬원활화단백격매(mitogen activated protein kinases,MAPKs)적아통로신호단백-세포외신호조절단백격매(extracellular signal-regulated kinase,ERK1/2)、P38화c-안기말단단백격매(c-Jun N-terminal kinase,JNK)재대서광기흡입폐손상조직중적표체급각통로재광기흡입성폐손상중적작용.방법 선취30지웅성Wistar대서수궤분성공기대조조、광기흡입조、PD98059(ERK1/2특이성조단제)、SB203580(P38특이성조단제)화SP600125(JNK특이성조단제)간예조,매조6지,응용정향류동광기흡입장치,복제대서광기(8.33 L/mg)흡입성폐손상적모형,공기대조조흡입공기,3개간예조균재광기(8.33 L/mg)흡입전급여PD98059、SB203580、SP600125.분별채용면역조화급단백질인적( Western blot)법검측폐조직MAPKs(ERK1/2、P38、JNK)단백급린산화단백p-MAPKs (p-ERK 1/2、p-P38、p-JNK)적정성정량표체.관찰폐조직병이학개변、계수지기관관세액(BALF)중중성립세포수、측정폐습간비.결과 공기대조조부폐포상피세포급기도상피세포견소량p-ERK1/2、p-P38、p-JNK양성표체；광기흡입조p-ERK1/2、p-P38、p-JNK양성세포수명현증다,엄범분포우폐내세포:폐포상피세포、기도상피세포、흉막간피세포、침윤염세포급간질섬유세포；간예조p-ERK1/2、p-P38、p-JNK양성세포수감소.각조ERK、P38、JNK적표체무명현변화；광기흡입조교공기대조조p-P38、p-JNK단백표체증강,3개간예조적p-ERK1/2、p-P38、p-JNK적단백표체수평균저우광기흡입조,차이균유통계학의의(P＜0.05,P＜0.01).여공기대조조[중성립세포계수:(2.05±0.75)×104,폐습간비:(3.64％±0.72％)]비교,광기흡입조폐손상정도엄중,표현출폐손상적전형병이학특정,차BALF중중성립세포수[(10.78±2.16)×104]증다,폐습간비(7.65％±0.58％)승고,SP600125간예조화SB203580간예조BALF중중성립세포수[SP600125조:(7.86±2.12)×104,SB203580조:(8.43±1.51)×104]화폐습간비[SP600125조:(6.10％±0.97％),SB203580조:(6.09％±1.43％)]강저,차이균유통계학의의(P＜0.05,P＜0.01).결론 광기흡입가능격활MAPK신호통로,통과p-MAPKs표체증가재광기흡입후적폐손상중발휘작용,기중이p-P38、p-JNK활성변화위주.
Objective This study aimed to investigate the expression and role of the mitogen activated protein kinases (ERK1/2,P38,JNK) in phosgene induced lung injury in rats in vivo.Method 30 male wistar rats were randomized into the group as follows,Gas inhalation control group,Phosgene inhalation group,and the following groups of the inhibitors of MAPK,involving SP600125,PD98059 and SB203580,6 animals in each group,we copy the model of phosgene-induced lung injury,used the directional flow-inhalation device,the air control group inhaled the air,and the intervention groups were given PD98059 (intraperitoneal injection),SB203580 (hypodermic injection),SP600125 (intravenous) respectively before the inhalation of phosgene.The locations and quantities of three subfamilies of MAPKs(ERK1/2,P38,JNK) and p-MAPKs (p-ERK1/2,p-P38,p-JNK) were analyzed by immunohistochemistry and Western Blot analysis respectively; The histopathological changes of lung tissues,the number of neutrophil cells and the W/D were examined.Result There were rare pERK1/2,p-P38 and p-JNK positive expression in alveolar and airway epithelial cells in control group,while the positive cells increased strikingly in phosgene inhalation groups,these ceils involved in this process mainly included alveolar epithelial cells,air way epithelial cells,pleuralmesothelial cells,infiltrative inflammatory cells,interstitium fibrocytes.After the intervention of the specific inhibitor,the positive cells decreased.As Western Blot analysis show,Protein quantities of p-P38 and pJNK were higher in phosgene inhalation groups than those in control group,and the differences were significant (P＜0.05).Protein quantities of p-ERK1/2,p-P38 and p-JNK were lower in intervention groups than phosgene inhalation group,and the differences were significant (P＜0.05,P＜0.01 ).The lung injury in phosgene inhalation groups was more severer compared with the control group,the typical pathological characters of acute lung injury were discovered,the increase of the number of neutrophil cells and W/D.After the intervention of the specific inhibitor SP600125 and SB203580,the number of neutrophil cells and W/D) reduced,and the differences were significant (P＜0.05,P＜0.01 ).Conclusion Phosgene inhalation may activate the MAPK signaling pathway,and the expression of the phosphorylation of MAPKs increased,especially the P38 ang JNK.The results may contribute to the lung injury induced by phosgene.
