中华危重病急救医学
中華危重病急救醫學
중화위중병급구의학
Chinese Critical Care Medicine
2015年
4期
280-284
,共5页
刘刚%宋冬梅%江宇%朱丽青%葛赟
劉剛%宋鼕梅%江宇%硃麗青%葛赟
류강%송동매%강우%주려청%갈빈
百草枯%中毒%肺损伤%血红素氧合酶-1
百草枯%中毒%肺損傷%血紅素氧閤酶-1
백초고%중독%폐손상%혈홍소양합매-1
Paraquat%Poisoning%Lung injury%Heme oxygenase-1
目的：观察百草枯中毒小鼠肺组织血红素氧合酶-1（HO-1）的表达，探讨其在百草枯中毒肺损伤中的机制。方法将58只雄性ICR小鼠按随机数字表法分为对照组（8只）和染毒组（50只），采用一次性灌胃20%百草枯溶液50 mg/kg建立中毒模型，对照组灌胃等量生理盐水。对照组于实验首日，染毒组于染毒后6 h及1、3、7、14 d分别处死8只存活小鼠取肺组织，行苏木素-伊红（HE）染色后光镜下观察肺组织病理改变；免疫组化法检测肺组织HO-1阳性表达；蛋白质免疫印迹试验（Western Blot）检测肺组织HO-1蛋白表达；检测超氧化物歧化酶（SOD）活性和丙二醛（MDA）含量。结果小鼠染毒后1 h即出现呼吸急促、乏力等症状，3～5 d时表现最为严重，14 d时饮食、活动恢复正常。光镜下观察显示，对照组肺组织无明显病理改变。染毒1 d时肺泡结构紊乱，肺组织出现明显的出血、水肿和炎性细胞浸润；3 d时肺组织病理改变加重；7 d时肺泡炎症表现减轻；14 d时炎症表现基本消失，但肺泡结构仍紊乱。免疫组化显示，对照组肺组织细胞中HO-1多不表达，少许轻度表达于气道上皮细胞。染毒1 d时HO-1主要分布于气道黏膜上皮细胞、炎性细胞、血管内皮细胞的胞质中；随后表达逐渐减少，7 d时恢复至对照组水平。染毒6 h时肺组织HO-1蛋白表达（灰度值）即较对照组明显升高（2.438±0.467比0.475±0.167，P＜0.01），1 d时达高峰（9.200±0.940比0.475±0.167，P＜0.01），持续至7 d时恢复至对照组水平（0.825±0.260比0.475±0.167，P＞0.05）。染毒6 h时肺组织SOD活性（μU/L）即较对照组明显降低（649.681±13.951比1167.051±15.744，P＜0.01），随后逐渐升高，至14 d仍显著低于对照组（859.733±121.079比1167.051±14.744，P＜0.01）；染毒6 h时肺组织MDA含量（μmol/L）即较对照组明显升高（4.542±0.266比3.705±0.176，P＜0.01），1 d时达到高峰（5.956±0.281比3.705±0.176，P＜0.01），随后逐渐下降，3 d时已恢复至对照组水平（4.134±0.168比3.705±0.176，P＞0.05）。结论急性百草枯中毒小鼠肺组织HO-1表达明显增高，可能是参与百草枯中毒肺损伤的重要保护机制。
目的：觀察百草枯中毒小鼠肺組織血紅素氧閤酶-1（HO-1）的錶達，探討其在百草枯中毒肺損傷中的機製。方法將58隻雄性ICR小鼠按隨機數字錶法分為對照組（8隻）和染毒組（50隻），採用一次性灌胃20%百草枯溶液50 mg/kg建立中毒模型，對照組灌胃等量生理鹽水。對照組于實驗首日，染毒組于染毒後6 h及1、3、7、14 d分彆處死8隻存活小鼠取肺組織，行囌木素-伊紅（HE）染色後光鏡下觀察肺組織病理改變；免疫組化法檢測肺組織HO-1暘性錶達；蛋白質免疫印跡試驗（Western Blot）檢測肺組織HO-1蛋白錶達；檢測超氧化物歧化酶（SOD）活性和丙二醛（MDA）含量。結果小鼠染毒後1 h即齣現呼吸急促、乏力等癥狀，3～5 d時錶現最為嚴重，14 d時飲食、活動恢複正常。光鏡下觀察顯示，對照組肺組織無明顯病理改變。染毒1 d時肺泡結構紊亂，肺組織齣現明顯的齣血、水腫和炎性細胞浸潤；3 d時肺組織病理改變加重；7 d時肺泡炎癥錶現減輕；14 d時炎癥錶現基本消失，但肺泡結構仍紊亂。免疫組化顯示，對照組肺組織細胞中HO-1多不錶達，少許輕度錶達于氣道上皮細胞。染毒1 d時HO-1主要分佈于氣道黏膜上皮細胞、炎性細胞、血管內皮細胞的胞質中；隨後錶達逐漸減少，7 d時恢複至對照組水平。染毒6 h時肺組織HO-1蛋白錶達（灰度值）即較對照組明顯升高（2.438±0.467比0.475±0.167，P＜0.01），1 d時達高峰（9.200±0.940比0.475±0.167，P＜0.01），持續至7 d時恢複至對照組水平（0.825±0.260比0.475±0.167，P＞0.05）。染毒6 h時肺組織SOD活性（μU/L）即較對照組明顯降低（649.681±13.951比1167.051±15.744，P＜0.01），隨後逐漸升高，至14 d仍顯著低于對照組（859.733±121.079比1167.051±14.744，P＜0.01）；染毒6 h時肺組織MDA含量（μmol/L）即較對照組明顯升高（4.542±0.266比3.705±0.176，P＜0.01），1 d時達到高峰（5.956±0.281比3.705±0.176，P＜0.01），隨後逐漸下降，3 d時已恢複至對照組水平（4.134±0.168比3.705±0.176，P＞0.05）。結論急性百草枯中毒小鼠肺組織HO-1錶達明顯增高，可能是參與百草枯中毒肺損傷的重要保護機製。
목적：관찰백초고중독소서폐조직혈홍소양합매-1（HO-1）적표체，탐토기재백초고중독폐손상중적궤제。방법장58지웅성ICR소서안수궤수자표법분위대조조（8지）화염독조（50지），채용일차성관위20%백초고용액50 mg/kg건립중독모형，대조조관위등량생리염수。대조조우실험수일，염독조우염독후6 h급1、3、7、14 d분별처사8지존활소서취폐조직，행소목소-이홍（HE）염색후광경하관찰폐조직병리개변；면역조화법검측폐조직HO-1양성표체；단백질면역인적시험（Western Blot）검측폐조직HO-1단백표체；검측초양화물기화매（SOD）활성화병이철（MDA）함량。결과소서염독후1 h즉출현호흡급촉、핍력등증상，3～5 d시표현최위엄중，14 d시음식、활동회복정상。광경하관찰현시，대조조폐조직무명현병리개변。염독1 d시폐포결구문란，폐조직출현명현적출혈、수종화염성세포침윤；3 d시폐조직병리개변가중；7 d시폐포염증표현감경；14 d시염증표현기본소실，단폐포결구잉문란。면역조화현시，대조조폐조직세포중HO-1다불표체，소허경도표체우기도상피세포。염독1 d시HO-1주요분포우기도점막상피세포、염성세포、혈관내피세포적포질중；수후표체축점감소，7 d시회복지대조조수평。염독6 h시폐조직HO-1단백표체（회도치）즉교대조조명현승고（2.438±0.467비0.475±0.167，P＜0.01），1 d시체고봉（9.200±0.940비0.475±0.167，P＜0.01），지속지7 d시회복지대조조수평（0.825±0.260비0.475±0.167，P＞0.05）。염독6 h시폐조직SOD활성（μU/L）즉교대조조명현강저（649.681±13.951비1167.051±15.744，P＜0.01），수후축점승고，지14 d잉현저저우대조조（859.733±121.079비1167.051±14.744，P＜0.01）；염독6 h시폐조직MDA함량（μmol/L）즉교대조조명현승고（4.542±0.266비3.705±0.176，P＜0.01），1 d시체도고봉（5.956±0.281비3.705±0.176，P＜0.01），수후축점하강，3 d시이회복지대조조수평（4.134±0.168비3.705±0.176，P＞0.05）。결론급성백초고중독소서폐조직HO-1표체명현증고，가능시삼여백초고중독폐손상적중요보호궤제。
ObjectiveTo investigate the expression of heme oxygenase-1 (HO-1) in lung tissue of mice with acute paraquat poisoning, and discuss its pathological mechanism.Methods Fifty-eight healthy male mice were randomly divided into control group (n = 8) and poisoned group (n = 50). The mice in poisoned group were lavaged with 20% paraquat (50 mg/kg), and those in control group with equal amount of normal saline. The mice were sacrificed on the day of experiment in control group, and those in poisoned group at 6 hours and 1, 3, 7, 14 days after poisoning. The lung tissue was harvested to observe the changes in pathology of lung with hematoxylin and eosin (HE) staining. The positive expression of HO-1 was determined with immunohistochemistry, and the protein expression of HO-1 was determined with Western Blot. The contents of superoxide dismutase (SOD) and malonaldehyde (MDA) were determined.Results The mice showed shortness of breath and signs of exhaustion 1 hour after poisoning, getting worse on 3-5 days, but returned to normal 14 days after poisoning. Under the light microscope, it showed that the control group had no significant pathological changes in lung tissue. One day after the ingestion, pulmonary alveolar structure disorder, obvious hemorrhage, edema and infiltration of inflammatory cells were found. At 3 days, the pathological changes in the lung tissue were more pronounced. They were less pronounced on 7 days, and inflammatory changes disappeared on 14th day, but alveolar structure disorder remained. Immunohistochemical test showed that HO-1 was seldom expressed in the lung tissue, and a little amount was expressed in the mucosal epithelial cells of the airway in control group. It was shown that inflammatory cell and endothelial were mainly distributed in the mucosal epithelial cells of airway 1 day after poisoning followed by a gradually decrease tendence, and came to normal level of control group 7 days after poisoning. It was shown by Western Blot that HO-1 (gray value) in lung tissue increased 6 hours after poisoning (2.438±0.467 vs. 0.475±0.167,P< 0.01), peaked at 1 day (9.200±0.940 vs. 0.475±0.167,P< 0.01), continued to increase till 7 days after poisoning, and it lowered to normal level thereafter (0.825±0.260 vs. 0.475±0.167,P> 0.05). The SOD activity (μU/L) in lung tissue was lowered 6 hours after poisoning, and it was significantly lower than that of control group (649.681±13.951 vs. 1 167.051±15.744,P< 0.01), and it continued to decrease up to 14 days after poisoning (859.733±121.079 vs. 1 167.051±14.744,P< 0.01). MDA content (μmol/L) in the lung tissue homogenate was elevated 6 hours after poisoning with significant difference compared with that of the control group (4.542±0.266 vs. 3.705±0.176,P< 0.01). It peaked on day 1 (5.956±0.281 vs. 3.705±0.176,P< 0.01), then it declined and reached normal level 3 days after poisoning (4.134±0.168 vs. 3.705±0.176,P> 0.05).Conclusion HO-1 expression was increased significantly in lung tissue of mice with acute paraquat poisoning, which may be considered as an important protection mechanism against paraquat poisoning.
