中华危重病急救医学
中華危重病急救醫學
중화위중병급구의학
Chinese Critical Care Medicine
2013年
6期
331-334
,共4页
孟潇潇%刘刊%谈玖婷%谢晖%王瑞兰
孟瀟瀟%劉刊%談玖婷%謝暉%王瑞蘭
맹소소%류간%담구정%사휘%왕서란
百草枯%中毒%内质网应激%氧化应激%肺纤维化
百草枯%中毒%內質網應激%氧化應激%肺纖維化
백초고%중독%내질망응격%양화응격%폐섬유화
Paraquat%Poisoning%Endoplasmic reticulum stress%Oxidative stress%Lung fibrosis
目的 探讨百草枯(PQ)中毒后大鼠发生肺纤维化与内质网应激的关系.方法 将100只雄性SD大鼠按随机数字表法分为对照组(10只)和染毒组(90只),20％PQ溶液灌胃染毒后2、6、12、24、48、72、96、168、336 h处死动物取肺组织,苏木素-伊红(HE)染色和Masson染色观察肺组织病理学变化；免疫组化检测糖调节蛋白78(GRP78)表达；测定肺组织丙二醛(MDA)含量；提取肺组织总蛋白,用蛋白质免疫印迹试验(Western blotting)检测α-平滑肌肌动蛋白(α-SMA)和GRP78的蛋白表达.结果 HE及Masson染色结果提示染毒组出现肺组织炎性细胞浸润和胶原纤维沉积,随中毒时间延长,肺纤维化逐渐加重.染毒后2 h MDA水平及α-SMA、GRP78蛋白表达均较对照组明显升高[MDA (nmol/mg):1.38±0.18比0.85±0.05,α-SMA:0.23±0.01比0.14±0.03,GRP78:0.72±0.02比0.37±0.06,P＜ 0.05或P＜0.01],随时间延长,MDA水平及α-SMA蛋白表达逐步升高,GRP78蛋白表达于染毒72 h后显著下降.结论 PQ中毒大鼠肺内质网应激标志性蛋白GRP78和成纤维细胞表型标志物α-SMA均增高,提示内质网应激在PQ中毒引起的肺纤维中发挥重要作用.
目的 探討百草枯(PQ)中毒後大鼠髮生肺纖維化與內質網應激的關繫.方法 將100隻雄性SD大鼠按隨機數字錶法分為對照組(10隻)和染毒組(90隻),20％PQ溶液灌胃染毒後2、6、12、24、48、72、96、168、336 h處死動物取肺組織,囌木素-伊紅(HE)染色和Masson染色觀察肺組織病理學變化；免疫組化檢測糖調節蛋白78(GRP78)錶達；測定肺組織丙二醛(MDA)含量；提取肺組織總蛋白,用蛋白質免疫印跡試驗(Western blotting)檢測α-平滑肌肌動蛋白(α-SMA)和GRP78的蛋白錶達.結果 HE及Masson染色結果提示染毒組齣現肺組織炎性細胞浸潤和膠原纖維沉積,隨中毒時間延長,肺纖維化逐漸加重.染毒後2 h MDA水平及α-SMA、GRP78蛋白錶達均較對照組明顯升高[MDA (nmol/mg):1.38±0.18比0.85±0.05,α-SMA:0.23±0.01比0.14±0.03,GRP78:0.72±0.02比0.37±0.06,P＜ 0.05或P＜0.01],隨時間延長,MDA水平及α-SMA蛋白錶達逐步升高,GRP78蛋白錶達于染毒72 h後顯著下降.結論 PQ中毒大鼠肺內質網應激標誌性蛋白GRP78和成纖維細胞錶型標誌物α-SMA均增高,提示內質網應激在PQ中毒引起的肺纖維中髮揮重要作用.
목적 탐토백초고(PQ)중독후대서발생폐섬유화여내질망응격적관계.방법 장100지웅성SD대서안수궤수자표법분위대조조(10지)화염독조(90지),20％PQ용액관위염독후2、6、12、24、48、72、96、168、336 h처사동물취폐조직,소목소-이홍(HE)염색화Masson염색관찰폐조직병이학변화；면역조화검측당조절단백78(GRP78)표체；측정폐조직병이철(MDA)함량；제취폐조직총단백,용단백질면역인적시험(Western blotting)검측α-평활기기동단백(α-SMA)화GRP78적단백표체.결과 HE급Masson염색결과제시염독조출현폐조직염성세포침윤화효원섬유침적,수중독시간연장,폐섬유화축점가중.염독후2 h MDA수평급α-SMA、GRP78단백표체균교대조조명현승고[MDA (nmol/mg):1.38±0.18비0.85±0.05,α-SMA:0.23±0.01비0.14±0.03,GRP78:0.72±0.02비0.37±0.06,P＜ 0.05혹P＜0.01],수시간연장,MDA수평급α-SMA단백표체축보승고,GRP78단백표체우염독72 h후현저하강.결론 PQ중독대서폐내질망응격표지성단백GRP78화성섬유세포표형표지물α-SMA균증고,제시내질망응격재PQ중독인기적폐섬유중발휘중요작용.
Objective To investigate the relationship between pulmonary fibrosis and endoplasmic reticulum stress (ERS) in rats with paraquat poisoning.Methods One hundred male Sprague-Dawley (SD) rats were randomly divided into control group (n =10) and paraquat poisoning group (n =90).The animals were sacrificed by exsanguination at 2,6,12,24,48,72,96,168,336 hours after administration with 20％ parquat solution.The paraffin sections of lung tissue were stained with hematoxylin-eosin (HE) and Masson trichrome to observe the pathological changes.Glucose-regulated protein 78 (GRP78) was determined by immunohistochemistry,and malondialdehyde (MDA) of lung tissue was measured.The total protein of tissue was abstracted,and the α-smooth muscle actin (α-SMA) and GRP78 was detected by Western blotting.Results HE and Masson staining demonstrated inflammatory infiltration and collagen deposition in the lung after paraquat administration with a tendency of exaggeration with time,and finally resulted in fibrosis.The expressions of MDA,α-SMA and GRP78 in the lung tissue were significantly increased 2 hours after paraquat administration compared with those of control group [MDA (nmol/mg):1.38 ±0.18 vs.0.85 ±0.05,α-SMA:0.23 ±0.01 vs.0.14 ±0.03,GRP78:0.72 ±0.02 vs.0.37 ±0.06,P＜0.05 or P＜0.01],and the expressions of MDA and α-SMA were gradually increased with time.GRP78 protein expression was decreased at 72 hours after paraquat administration.Conclusion The results reveal that the expressions of α-SMA and GRP78 in paraquat poisoning group are up-regulated,suggesting ERS plays an important role in paraquat induced-pulmonary fibrosis.
