中华急诊医学杂志
中華急診醫學雜誌
중화급진의학잡지
CHINESE JOURNAL OF EMERGENCY MEDICINE
2012年
2期
164-170
,共7页
葛赟%余毅娟%郑嘉奕%刘瑶%邱俏檬%洪广亮%胡国新%李萌芳%卢中秋
葛赟%餘毅娟%鄭嘉奕%劉瑤%邱俏檬%洪廣亮%鬍國新%李萌芳%盧中鞦
갈빈%여의연%정가혁%류요%구초몽%홍엄량%호국신%리맹방%로중추
硫化氢%乌司他丁%急性肺损伤%氧化应激%核转录因子红系相关因子-2%血红素氧合酶-1%醌氧化还原酶-1%大鼠
硫化氫%烏司他丁%急性肺損傷%氧化應激%覈轉錄因子紅繫相關因子-2%血紅素氧閤酶-1%醌氧化還原酶-1%大鼠
류화경%오사타정%급성폐손상%양화응격%핵전록인자홍계상관인자-2%혈홍소양합매-1%곤양화환원매-1%대서
Hydrogen sulfide%Ulinastratin%Acute lung injury%Oxidative Stress%Nuclear factor-E2-related factor2%Heme oxygenase 1%NAD(P)H: quinine oxidoreductase 1%Rats
目的 观察硫化氢(hydrogen sulfide,H2S)急性中毒大鼠肺组织血红素氧合酶-1(heme oxygenase 1,HO-1)、醌氧化还原酶-1(NAD(P)H:quinine oxidoreductase 1,NQO-1)和核转录因子红系相关因子-2(nuclear factor E2-related factor 2,Nrf2)的动态变化及乌司他丁(ulinastatin,UTI对其的影响.方法 将清洁级SD大鼠96只随机(随机数字法)分为健康对照组(NS组,n=8),UTI对照组(UTI组,n=8),H2S染毒模型组(H2S组,n=40,采用在染毒柜内暴露吸入体积分数200×10-6的H2S1h,构建H2S急性染毒模型)和UTI干预组(H2S+UTI组,n=40,建立模型后立即腹腔注射UTI 10万U/kg).后两组大鼠于染毒后2、6、12、24、48 h时点麻醉后活杀,留取肺标本.观察大鼠行为学变化,ELISA法测定肺组织中HO-1和NQO-1活力的动态变化；RT-PCR法检测肺组织HO-1、NQO-1和Nrf2 mRNA表达,Western blot法检测Nrf2蛋白表达.观察肺组织病理学改变并行肺损伤评分.结果 与NS组比较,H2S组在染毒后2、6、12h时点HO-1活力和mRNA表达均明显升高(P＜0.01),其中2h呈高峰；与同时间点H2S组比较,H2S+ UTI组染毒后6、12、24、48 h时点HO-1活力和mRNA表达显著升高(P＜0.01).与 、NS组比较,H2S组在染毒后2、6、12、24 h时点NQO-1活力和mRNA表达均明显升高(P＜0.01)其中2h呈高峰；与同时间点H2S组比较,H2S+UTI组染毒后6、12、24、48 h时点NQO-1活力和mRNA表达显著升高(P＜0.01).与NS组比较,H2S组在染毒后2、6、12 h时点Nrf2mRNA和蛋白表达均明显升高(P＜0.01或P＜0.05),其中2h呈高峰；与同时间点H2S组比较,H2S+UTI组染毒后6、12、24、48 h时点Nrf2 mRNA和蛋白表达显著升高(P＜0.01).光镜下,H2S组在染毒后24 h大鼠肺组织损害明显；H2S+UTI组大鼠肺损伤较H2S组有所减轻.在染毒后12、24、48 h时点H2S+UTI组的肺损伤评分明显低于H2S组(P＜0.01).结论 HO-1、NQO-1和Nrf2参与了H2S中毒急性肺损伤的病理生理学过程；UTI干预治疗能显著提高HO-1、NQO-1及Nrf2的活力及表达,减轻H2S中毒急性肺损伤.
目的 觀察硫化氫(hydrogen sulfide,H2S)急性中毒大鼠肺組織血紅素氧閤酶-1(heme oxygenase 1,HO-1)、醌氧化還原酶-1(NAD(P)H:quinine oxidoreductase 1,NQO-1)和覈轉錄因子紅繫相關因子-2(nuclear factor E2-related factor 2,Nrf2)的動態變化及烏司他丁(ulinastatin,UTI對其的影響.方法 將清潔級SD大鼠96隻隨機(隨機數字法)分為健康對照組(NS組,n=8),UTI對照組(UTI組,n=8),H2S染毒模型組(H2S組,n=40,採用在染毒櫃內暴露吸入體積分數200×10-6的H2S1h,構建H2S急性染毒模型)和UTI榦預組(H2S+UTI組,n=40,建立模型後立即腹腔註射UTI 10萬U/kg).後兩組大鼠于染毒後2、6、12、24、48 h時點痳醉後活殺,留取肺標本.觀察大鼠行為學變化,ELISA法測定肺組織中HO-1和NQO-1活力的動態變化；RT-PCR法檢測肺組織HO-1、NQO-1和Nrf2 mRNA錶達,Western blot法檢測Nrf2蛋白錶達.觀察肺組織病理學改變併行肺損傷評分.結果 與NS組比較,H2S組在染毒後2、6、12h時點HO-1活力和mRNA錶達均明顯升高(P＜0.01),其中2h呈高峰；與同時間點H2S組比較,H2S+ UTI組染毒後6、12、24、48 h時點HO-1活力和mRNA錶達顯著升高(P＜0.01).與 、NS組比較,H2S組在染毒後2、6、12、24 h時點NQO-1活力和mRNA錶達均明顯升高(P＜0.01)其中2h呈高峰；與同時間點H2S組比較,H2S+UTI組染毒後6、12、24、48 h時點NQO-1活力和mRNA錶達顯著升高(P＜0.01).與NS組比較,H2S組在染毒後2、6、12 h時點Nrf2mRNA和蛋白錶達均明顯升高(P＜0.01或P＜0.05),其中2h呈高峰；與同時間點H2S組比較,H2S+UTI組染毒後6、12、24、48 h時點Nrf2 mRNA和蛋白錶達顯著升高(P＜0.01).光鏡下,H2S組在染毒後24 h大鼠肺組織損害明顯；H2S+UTI組大鼠肺損傷較H2S組有所減輕.在染毒後12、24、48 h時點H2S+UTI組的肺損傷評分明顯低于H2S組(P＜0.01).結論 HO-1、NQO-1和Nrf2參與瞭H2S中毒急性肺損傷的病理生理學過程；UTI榦預治療能顯著提高HO-1、NQO-1及Nrf2的活力及錶達,減輕H2S中毒急性肺損傷.
목적 관찰류화경(hydrogen sulfide,H2S)급성중독대서폐조직혈홍소양합매-1(heme oxygenase 1,HO-1)、곤양화환원매-1(NAD(P)H:quinine oxidoreductase 1,NQO-1)화핵전록인자홍계상관인자-2(nuclear factor E2-related factor 2,Nrf2)적동태변화급오사타정(ulinastatin,UTI대기적영향.방법 장청길급SD대서96지수궤(수궤수자법)분위건강대조조(NS조,n=8),UTI대조조(UTI조,n=8),H2S염독모형조(H2S조,n=40,채용재염독거내폭로흡입체적분수200×10-6적H2S1h,구건H2S급성염독모형)화UTI간예조(H2S+UTI조,n=40,건립모형후립즉복강주사UTI 10만U/kg).후량조대서우염독후2、6、12、24、48 h시점마취후활살,류취폐표본.관찰대서행위학변화,ELISA법측정폐조직중HO-1화NQO-1활력적동태변화；RT-PCR법검측폐조직HO-1、NQO-1화Nrf2 mRNA표체,Western blot법검측Nrf2단백표체.관찰폐조직병이학개변병행폐손상평분.결과 여NS조비교,H2S조재염독후2、6、12h시점HO-1활력화mRNA표체균명현승고(P＜0.01),기중2h정고봉；여동시간점H2S조비교,H2S+ UTI조염독후6、12、24、48 h시점HO-1활력화mRNA표체현저승고(P＜0.01).여 、NS조비교,H2S조재염독후2、6、12、24 h시점NQO-1활력화mRNA표체균명현승고(P＜0.01)기중2h정고봉；여동시간점H2S조비교,H2S+UTI조염독후6、12、24、48 h시점NQO-1활력화mRNA표체현저승고(P＜0.01).여NS조비교,H2S조재염독후2、6、12 h시점Nrf2mRNA화단백표체균명현승고(P＜0.01혹P＜0.05),기중2h정고봉；여동시간점H2S조비교,H2S+UTI조염독후6、12、24、48 h시점Nrf2 mRNA화단백표체현저승고(P＜0.01).광경하,H2S조재염독후24 h대서폐조직손해명현；H2S+UTI조대서폐손상교H2S조유소감경.재염독후12、24、48 h시점H2S+UTI조적폐손상평분명현저우H2S조(P＜0.01).결론 HO-1、NQO-1화Nrf2삼여료H2S중독급성폐손상적병리생이학과정；UTI간예치료능현저제고HO-1、NQO-1급Nrf2적활력급표체,감경H2S중독급성폐손상.
Objective To observe the dynamic changes of heme oxygenase 1,NAD(P)H:quinine oxidoreductase 1 and Nuclear factor-E2-related factor 2 in the lung tissue of acute H2S-intoxicated rats and intervention effects of ulinastratin(UTI).Methods A total of 96 SD rats of clean grade were divided randomly(random number)into four groups:normal control group(NS group,n =8),UTI control group(UTI group,n =8),H2S-intoxicated model group(H2S group,n =40,rats were exposed to H2S(200 × 10-6)for 1 h to establish the H2S-intoxicated model)and UTI treatment group(H2S +UTI group,n =40,rats were intraperitoneal injected with the dose of UTI 105 U/kg).H2S group and H2S + UTI group were sacrificed 2,6,12,24 and 48 h after modeling.The activity and mRNA expression of HO-1 and NQO-1 in the lung tissue were measured by ELISA and RT-PCR methods,and the expression of Nrf2 mRNA and protein in the lung tissue was detected by RT-PCR and Western Blot methods.Pathological changes of lung tissue were observed by lightmicroscope and the lung injury score was used to evaluate inhalation injury.Results The pulmonary HO-1 activity and mRNA expression in rats of H2S group at 2,6,12 h(P ＜ 0.01)after intoxication were markedly increased than that in NS group:In comparison with H2S group,the pulmonary HO-1 activity and mRNA expression increased at 6,12,24,48 h(P ＜0.01).The pulmonary NQO-1 activity and mRNA expression in rats of H2S group at 2,6,12,24 h(P＜ 0.01)after intoxication were markedly increased than that in NS group; In comparison with H2S group,the pulmonary NQO-1 activity and mRNA expression increased at 6,12,24,48 h(P ＜ 0.01).The pulmonary Nrf2 mRNA and protein expression in rats of H2S group at 2,6,12 h(P ＜0.01 or P ＜0.05)after modeling were markedly increased than that in NS group and reached peak 2 hour after modeling; In comparison with H2S group,the pulmonary Nrf2 mRNA and protein expression increased at 6,12,24,48 h(P ＜0.01).At 24 h after modeling,the degree of lung damage were also decreased in H2S group compared with H2S + UTI group in the lightmicroscope.Histopathological examination showed that the degree of lung injury in H2S + UTI group was less severe than that in H2S group especially in the 12,24 and 48 h (P ＜0.01).Conclusions HO-1,NQO-1 and Nrf2 are involved in the pathogenesis of acute lung injury induced by H2S-intoxicated in rats.UTI may improve the imbalance in redox and activate HO-1,NQO-1 and Nrf2 can reduce lung injury and protect the lung injury induced by H2S in rats.
