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
274-279
,共6页
曹玉芳%李景辉%欧宗兴%殷宗宝%陈栩栩%韩艳丽%宋维
曹玉芳%李景輝%歐宗興%慇宗寶%陳栩栩%韓豔麗%宋維
조옥방%리경휘%구종흥%은종보%진허허%한염려%송유
百草枯%中毒%核转录因子-κB%肿瘤坏死因子-α%急性肺损伤%二烯丙基硫化物
百草枯%中毒%覈轉錄因子-κB%腫瘤壞死因子-α%急性肺損傷%二烯丙基硫化物
백초고%중독%핵전록인자-κB%종류배사인자-α%급성폐손상%이희병기류화물
Paraquat%Poisoning%Nuclear factor-κB%Tumor necrosis factor-α%Acute lung injury%Diallyl sulfide
目的：探讨二烯丙基硫化物（DAS）对百草枯中毒所致大鼠急性肺损伤（ALI）的器官保护作用及抗炎机制。方法将80只雄性Wistar大鼠按随机数字表法分为对照组、模型组、地塞米松（DXM）治疗组及DAS治疗组，每组20只。一次性灌胃百草枯溶液70 mg/kg制备中毒模型；对照组灌胃等量生理盐水。制模后当日分别经腹腔注射DAS 100 mg/kg（DAS治疗组）、等量生理盐水（模型组）或DXM 1 mg/kg（DXM治疗组），均每日1次，共14 d。各组分别于制模后1、3、7、14 d随机处死5只大鼠，取右肺下叶行苏木素-伊红（HE）染色，光镜下观察肺损伤程度；取右肺上叶计算湿/干质量（W/D）比值，评价肺水肿程度；取右肺中叶,采用免疫组化检测核转录因子-κB（NF-κB）表达；取左肺采用反转录-聚合酶链反应（RT-PCR）检测肿瘤坏死因子-α（TNF-α）mRNA表达。结果① HE染色显示：对照组肺组织肺泡结构完整；模型组病理改变逐渐加重，肺泡壁结构破坏、肺间质水肿及大量巨噬细胞、中性粒细胞浸润，并伴有炎细胞移行至肺泡腔，部分肺泡腔内形成透明膜； DAS治疗组和DXM治疗组肺间质有部分增生、少量炎性细胞浸润，肺泡结构均无明显破坏。②肺W/D比值：模型组各时间点肺W/D比值明显高于对照组，3 d时达最高值（6.15±0.54比4.15±2.10，P＜0.05）；DAS治疗组和DXM治疗组各时间点肺W/D比值明显低于模型组，以3 d为明显（3.99±1.26、4.30±0.70比6.15±0.54，均P＜0.05）；而DAS治疗组与DXM治疗组比较均无明显差异。③免疫组化显示：对照组细胞质有极少量NF-κBp65阳性表达；模型组细胞核中有较多NF-κBp65阳性表达；DAS治疗组和DXM治疗组细胞质中有少量NF-κBp65阳性表达，细胞核中无明显NF-κBp65阳性表达。积分A值明显低于模型组，以3 d为明显〔（17.98±0.06）×107、（18.53±0.04）×107比（28.85±0.61）×107，均P＜0.01〕；而DAS治疗组与DXM治疗组比较均无明显差异。④ RT-PCR显示：3 d时模型组肺组织TNF-α mRNA表达明显高于对照组（灰度值：3.63±0.62比0.51±0.13，P＜0.05）；DAS治疗组和DXM治疗组肺组织TNF-α mRNA表达较模型组明显下降（灰度值：2.49±0.57、2.02±0.26比3.63±0.62，均P＜0.05），而DAS治疗组与DXM治疗组比较无明显差异。结论腹腔注射DAS能降低百草枯诱导大鼠ALI时的肺部毛细血管通透性增加，减轻肺水肿，抑制肺组织中NF-κB、TNF-α表达，改善肺组织炎症病理改变。
目的：探討二烯丙基硫化物（DAS）對百草枯中毒所緻大鼠急性肺損傷（ALI）的器官保護作用及抗炎機製。方法將80隻雄性Wistar大鼠按隨機數字錶法分為對照組、模型組、地塞米鬆（DXM）治療組及DAS治療組，每組20隻。一次性灌胃百草枯溶液70 mg/kg製備中毒模型；對照組灌胃等量生理鹽水。製模後噹日分彆經腹腔註射DAS 100 mg/kg（DAS治療組）、等量生理鹽水（模型組）或DXM 1 mg/kg（DXM治療組），均每日1次，共14 d。各組分彆于製模後1、3、7、14 d隨機處死5隻大鼠，取右肺下葉行囌木素-伊紅（HE）染色，光鏡下觀察肺損傷程度；取右肺上葉計算濕/榦質量（W/D）比值，評價肺水腫程度；取右肺中葉,採用免疫組化檢測覈轉錄因子-κB（NF-κB）錶達；取左肺採用反轉錄-聚閤酶鏈反應（RT-PCR）檢測腫瘤壞死因子-α（TNF-α）mRNA錶達。結果① HE染色顯示：對照組肺組織肺泡結構完整；模型組病理改變逐漸加重，肺泡壁結構破壞、肺間質水腫及大量巨噬細胞、中性粒細胞浸潤，併伴有炎細胞移行至肺泡腔，部分肺泡腔內形成透明膜； DAS治療組和DXM治療組肺間質有部分增生、少量炎性細胞浸潤，肺泡結構均無明顯破壞。②肺W/D比值：模型組各時間點肺W/D比值明顯高于對照組，3 d時達最高值（6.15±0.54比4.15±2.10，P＜0.05）；DAS治療組和DXM治療組各時間點肺W/D比值明顯低于模型組，以3 d為明顯（3.99±1.26、4.30±0.70比6.15±0.54，均P＜0.05）；而DAS治療組與DXM治療組比較均無明顯差異。③免疫組化顯示：對照組細胞質有極少量NF-κBp65暘性錶達；模型組細胞覈中有較多NF-κBp65暘性錶達；DAS治療組和DXM治療組細胞質中有少量NF-κBp65暘性錶達，細胞覈中無明顯NF-κBp65暘性錶達。積分A值明顯低于模型組，以3 d為明顯〔（17.98±0.06）×107、（18.53±0.04）×107比（28.85±0.61）×107，均P＜0.01〕；而DAS治療組與DXM治療組比較均無明顯差異。④ RT-PCR顯示：3 d時模型組肺組織TNF-α mRNA錶達明顯高于對照組（灰度值：3.63±0.62比0.51±0.13，P＜0.05）；DAS治療組和DXM治療組肺組織TNF-α mRNA錶達較模型組明顯下降（灰度值：2.49±0.57、2.02±0.26比3.63±0.62，均P＜0.05），而DAS治療組與DXM治療組比較無明顯差異。結論腹腔註射DAS能降低百草枯誘導大鼠ALI時的肺部毛細血管通透性增加，減輕肺水腫，抑製肺組織中NF-κB、TNF-α錶達，改善肺組織炎癥病理改變。
목적：탐토이희병기류화물（DAS）대백초고중독소치대서급성폐손상（ALI）적기관보호작용급항염궤제。방법장80지웅성Wistar대서안수궤수자표법분위대조조、모형조、지새미송（DXM）치료조급DAS치료조，매조20지。일차성관위백초고용액70 mg/kg제비중독모형；대조조관위등량생리염수。제모후당일분별경복강주사DAS 100 mg/kg（DAS치료조）、등량생리염수（모형조）혹DXM 1 mg/kg（DXM치료조），균매일1차，공14 d。각조분별우제모후1、3、7、14 d수궤처사5지대서，취우폐하협행소목소-이홍（HE）염색，광경하관찰폐손상정도；취우폐상협계산습/간질량（W/D）비치，평개폐수종정도；취우폐중협,채용면역조화검측핵전록인자-κB（NF-κB）표체；취좌폐채용반전록-취합매련반응（RT-PCR）검측종류배사인자-α（TNF-α）mRNA표체。결과① HE염색현시：대조조폐조직폐포결구완정；모형조병리개변축점가중，폐포벽결구파배、폐간질수종급대량거서세포、중성립세포침윤，병반유염세포이행지폐포강，부분폐포강내형성투명막； DAS치료조화DXM치료조폐간질유부분증생、소량염성세포침윤，폐포결구균무명현파배。②폐W/D비치：모형조각시간점폐W/D비치명현고우대조조，3 d시체최고치（6.15±0.54비4.15±2.10，P＜0.05）；DAS치료조화DXM치료조각시간점폐W/D비치명현저우모형조，이3 d위명현（3.99±1.26、4.30±0.70비6.15±0.54，균P＜0.05）；이DAS치료조여DXM치료조비교균무명현차이。③면역조화현시：대조조세포질유겁소량NF-κBp65양성표체；모형조세포핵중유교다NF-κBp65양성표체；DAS치료조화DXM치료조세포질중유소량NF-κBp65양성표체，세포핵중무명현NF-κBp65양성표체。적분A치명현저우모형조，이3 d위명현〔（17.98±0.06）×107、（18.53±0.04）×107비（28.85±0.61）×107，균P＜0.01〕；이DAS치료조여DXM치료조비교균무명현차이。④ RT-PCR현시：3 d시모형조폐조직TNF-α mRNA표체명현고우대조조（회도치：3.63±0.62비0.51±0.13，P＜0.05）；DAS치료조화DXM치료조폐조직TNF-α mRNA표체교모형조명현하강（회도치：2.49±0.57、2.02±0.26비3.63±0.62，균P＜0.05），이DAS치료조여DXM치료조비교무명현차이。결론복강주사DAS능강저백초고유도대서ALI시적폐부모세혈관통투성증가，감경폐수종，억제폐조직중NF-κB、TNF-α표체，개선폐조직염증병리개변。
ObjectiveTo investigate the mechanism of anti-inflammatory effect of diallyl sulfide (DAS) in protection against acute lung injury (ALI) in rats with paraquat poisoning.Methods Eighty male Wistar rats were randomly divided into four groups, namely: control group, model group, dexamethasone (DXM) treatment group, and DAS treatment group, with 20 rats in each group. The model of paraquat poisoning was reproduced by single does of 70 mg/kg given by gavage, while the same volume of normal saline (NS) was given in same manner in control group. 100 mg/kg of DAS, the same volume of NS, or 1 mg/kg DXM injection were given respectively in DAS treatment group, model group, or DXM treatment group intraperitoneally after exposure to paraquat, once a day for 14 days. Five rats in each group were sacrificed at 1, 3, 7, 14 days, respectively. The inferior lobe of right lung was harvested, and the degree of lung injury was observed with hematoxylin and eosin (HE) staining under optical microscope; the upper lobe of right lung was used to determine the lung wet/dry weight (W/D) ratio and for evaluation of the degree of pulmonary edema. The expression of nuclear factor -κB (NF-κB) in the middle lobe of right lung was assessed with immunohistochemistry. The expression of tumor necrosis factor -α (TNF-α) mRNA in the left lung was determined with the reverse transcription-polymerase chain reaction (RT-PCR).Results① The pulmonary structure in control group was found to be intact. However, in the model group there were progressive pathological changes in lung, including marked edema and thickening of alveolar walls, collapse of alveoli, infiltration of inflammatory cells, alveolar wall, and obvious bleeding in the local lung tissue, and formation of transparent membrane in alveolar space. Less infiltration of inflammatory cells and no obvious destruction were found in alveolar structure in the DAS and DXM treatment groups.② Lung W/D ratio: lung W/D ratio of model group was apparently higher than that in control group at every time point, and peaking on the 3rd day (6.15±0.54 vs. 4.15±2.10,P< 0.05), and the ratio of lung W/D of DAS and DXM treatment groups was obviously lower than that in model group at every time point, especially on the 3rd day (3.99±1.26, 4.30±0.70 vs. 6.15±0.54, bothP< 0.05), but there was no significant difference between DAS and DXM treatment groups in this regard.③ The immunocytochemistry analysis revealed minimal NF-κBp65 expression in the cell nuclei of the control group, while extensive NF-κBp65 expression was found in model group. Minimal NF-κBp65 positive expression in the cytoplasm and even less positive expression in the nucleus was found in the DAS and DXM treatment groups, and integralA value was significantly lower in the DAS and DXM treatment groups than that of the model group, especially on the 3rd day [(17.98±0.06)×107, (18.53±0.04)×107 vs. (28.85±0.61)×107, bothP< 0.01], but there was no significant difference between DAS and DXM treatment groups.④ It was shown by RT-PCR that the expression of TNF-α mRNA in lung tissue of the model group was significantly higher than that in the control group on the 3rd day (gray value: 3.63±0.62 vs. 0.51±0.13, P< 0.05). The expression of TNF-α mRNA in lung tissue was significantly decreased in DAS and DXM treatment groups compared with model group (gray value: 2.49±0.57, 2.02±0.26 vs. 3.63±0.62, bothP< 0.05), but there was no significant difference between DAS and DXM treated groups.ConclusionTreatment with an intraperitoneally injection of DAS is capable of attenuate the extent of PQ-induced ALI in rats by alleviating pulmonary edema, inhibiting the expression of NF-κB and TNF-α in lung tissue, and ameliorating pathological changes in lung tissue.
