国际麻醉学与复苏杂志
國際痳醉學與複囌雜誌
국제마취학여복소잡지
INTERNATIONAL JOURNAL OF ANESTHESIOLOGY AND RESUSCITATION
2014年
4期
324-330,345
,共8页
细胞外调节蛋白激酶%核转录因子-κB%急性肺损伤%肿瘤坏死因子-α%白介素-10%诱导型一氧化氮合酶%内毒素
細胞外調節蛋白激酶%覈轉錄因子-κB%急性肺損傷%腫瘤壞死因子-α%白介素-10%誘導型一氧化氮閤酶%內毒素
세포외조절단백격매%핵전록인자-κB%급성폐손상%종류배사인자-α%백개소-10%유도형일양화담합매%내독소
Extracellular signal-regulated kinases%Nuclear factor-κB%Acute lung injury%Tumor necrosis factor-α%Interleukin-10%Inducible nitric oxide synthase%Lipopolysaccharide
目的 探讨细胞外调节蛋白激酶(extracellular signal-regulated kinases,ERK)信号通路在大鼠内毒素(lipopolysaccharide,LPS)性急性肺损伤(acute lung injury,ALI)中的作用. 方法 120只健康成年雄性SD大鼠按随机数字表法随机分为4组(每组30只):生理盐水组(Saline组),LPS组,LPS+U0126组(LPS+U组),生理盐水+U0126组(Saline+U组).其中各组又按照不同的时间点被分为1、3、6、12、24h5个亚组.参照文献及预实验,LPS组采用腹腔注射LPS 7.5 mg/kg(5 g/L)制作LPS性ALI模型,Saline组注射等体积的生理盐水.LPS+U组和Saline+U组在注射LPS或生理盐水前30 min,经腹腔注射U0126 5 mg/kg.利用Western blot检测大鼠肺组织中的磷酸化的ERK、高迁移率族蛋白B1(high-mobility group box 1,HMGB-1),胞核内的核转录因子-κB(nuclear factor-κB,NF-κB)p65的表达.同时检测支气管肺泡灌洗液(bronchoalveolar lavage fluid,BALF)中肿瘤坏死因子-α(tumor necrosis factor,TNF-α)、诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)和白介素(interleukin,IL)-10的浓度,肺脏水含量以及肺组织病理学的变化. 结果 注射LPS后,大鼠肺组织中磷酸化的ERK、HMGB-1明显增多,同时核内NF-κBp65的表达在3h和12h明显升高.BALF中的TNF-α、iNOS和IL-10的浓度也随之升高,肺水含量上升,组织病理学检查显示:肺组织出现明显的病理损害.使用U0126能明显抑制ERK的磷酸化、NF-κB的活化及HMGB-1的表达,肺水含量下降[(4.59±0.51) vs (5.19±0.10),P＜0.05)],BALF中TNF-α、iNOS和IL-10的浓度显著降低[TNF-α,3h达高峰,(28±3)vs(70±10),P＜0.05;iNOS,12h达高峰,(7 771±957) vs (10 679±1641),P＜0.05;IL-10,6h达高峰,(59±10)vs (91±11),P＜0.05],同时肺组织的病理损害也明显减轻. 结论 LPS性ALI过程中ERK活化,其抑制剂U0126能减轻LPS性ALI程度;ERK-NF-κB信号通路参与调控炎性因子的释放.
目的 探討細胞外調節蛋白激酶(extracellular signal-regulated kinases,ERK)信號通路在大鼠內毒素(lipopolysaccharide,LPS)性急性肺損傷(acute lung injury,ALI)中的作用. 方法 120隻健康成年雄性SD大鼠按隨機數字錶法隨機分為4組(每組30隻):生理鹽水組(Saline組),LPS組,LPS+U0126組(LPS+U組),生理鹽水+U0126組(Saline+U組).其中各組又按照不同的時間點被分為1、3、6、12、24h5箇亞組.參照文獻及預實驗,LPS組採用腹腔註射LPS 7.5 mg/kg(5 g/L)製作LPS性ALI模型,Saline組註射等體積的生理鹽水.LPS+U組和Saline+U組在註射LPS或生理鹽水前30 min,經腹腔註射U0126 5 mg/kg.利用Western blot檢測大鼠肺組織中的燐痠化的ERK、高遷移率族蛋白B1(high-mobility group box 1,HMGB-1),胞覈內的覈轉錄因子-κB(nuclear factor-κB,NF-κB)p65的錶達.同時檢測支氣管肺泡灌洗液(bronchoalveolar lavage fluid,BALF)中腫瘤壞死因子-α(tumor necrosis factor,TNF-α)、誘導型一氧化氮閤酶(inducible nitric oxide synthase,iNOS)和白介素(interleukin,IL)-10的濃度,肺髒水含量以及肺組織病理學的變化. 結果 註射LPS後,大鼠肺組織中燐痠化的ERK、HMGB-1明顯增多,同時覈內NF-κBp65的錶達在3h和12h明顯升高.BALF中的TNF-α、iNOS和IL-10的濃度也隨之升高,肺水含量上升,組織病理學檢查顯示:肺組織齣現明顯的病理損害.使用U0126能明顯抑製ERK的燐痠化、NF-κB的活化及HMGB-1的錶達,肺水含量下降[(4.59±0.51) vs (5.19±0.10),P＜0.05)],BALF中TNF-α、iNOS和IL-10的濃度顯著降低[TNF-α,3h達高峰,(28±3)vs(70±10),P＜0.05;iNOS,12h達高峰,(7 771±957) vs (10 679±1641),P＜0.05;IL-10,6h達高峰,(59±10)vs (91±11),P＜0.05],同時肺組織的病理損害也明顯減輕. 結論 LPS性ALI過程中ERK活化,其抑製劑U0126能減輕LPS性ALI程度;ERK-NF-κB信號通路參與調控炎性因子的釋放.
목적 탐토세포외조절단백격매(extracellular signal-regulated kinases,ERK)신호통로재대서내독소(lipopolysaccharide,LPS)성급성폐손상(acute lung injury,ALI)중적작용. 방법 120지건강성년웅성SD대서안수궤수자표법수궤분위4조(매조30지):생리염수조(Saline조),LPS조,LPS+U0126조(LPS+U조),생리염수+U0126조(Saline+U조).기중각조우안조불동적시간점피분위1、3、6、12、24h5개아조.삼조문헌급예실험,LPS조채용복강주사LPS 7.5 mg/kg(5 g/L)제작LPS성ALI모형,Saline조주사등체적적생리염수.LPS+U조화Saline+U조재주사LPS혹생리염수전30 min,경복강주사U0126 5 mg/kg.이용Western blot검측대서폐조직중적린산화적ERK、고천이솔족단백B1(high-mobility group box 1,HMGB-1),포핵내적핵전록인자-κB(nuclear factor-κB,NF-κB)p65적표체.동시검측지기관폐포관세액(bronchoalveolar lavage fluid,BALF)중종류배사인자-α(tumor necrosis factor,TNF-α)、유도형일양화담합매(inducible nitric oxide synthase,iNOS)화백개소(interleukin,IL)-10적농도,폐장수함량이급폐조직병이학적변화. 결과 주사LPS후,대서폐조직중린산화적ERK、HMGB-1명현증다,동시핵내NF-κBp65적표체재3h화12h명현승고.BALF중적TNF-α、iNOS화IL-10적농도야수지승고,폐수함량상승,조직병이학검사현시:폐조직출현명현적병리손해.사용U0126능명현억제ERK적린산화、NF-κB적활화급HMGB-1적표체,폐수함량하강[(4.59±0.51) vs (5.19±0.10),P＜0.05)],BALF중TNF-α、iNOS화IL-10적농도현저강저[TNF-α,3h체고봉,(28±3)vs(70±10),P＜0.05;iNOS,12h체고봉,(7 771±957) vs (10 679±1641),P＜0.05;IL-10,6h체고봉,(59±10)vs (91±11),P＜0.05],동시폐조직적병리손해야명현감경. 결론 LPS성ALI과정중ERK활화,기억제제U0126능감경LPS성ALI정도;ERK-NF-κB신호통로삼여조공염성인자적석방.
Objective To investigate the role of extracellular signal-regulated kinases (ERK) signal pathway in the lipopolysaccharide(LPS)-induced acute lung injury(ALI) in rats.Methods One hundred and twenty male SD rats were randomly divided into four groups (n=30):Saline group,LPS group,LSP+U group and Saline+U group.Each group was divided into five subgroups at 1,3,6,12 h and 24 h time point respectively.Western blot was used to detect the expression of phosphorylation of ERK1/2 and high-mobility group box 1 (HMGB-1) in lung tissues,nuclear factor-κB (NF-κB)p65 in the nuclear extracts,which reflected the extent of lung injury.Additionally we examined the concentration of tumor necrosis factor(TNF)-α,interleukin(IL)-10and inducible nitric oxide synthase (iNOS) in bronchoalveolar lavage fluid (BALF),the lung water content and the histopathologic changes of lung.In addition,survival rate was investigated between LPS group and LSP+U group,and each group with other 10 rats.Results With the administration of LPS,the phospho-ERK1/2 substantially increased immediately and subsequently the expression of NF-κB in the nuclear extracts was increased significantly and reached its peak level at 3 h and 12 h.Moreover,HMGB-1,one of the key mediators in the development of sepsis increased significantly after LPS administration.The concentrations of TNF-α,iNOS and IL-10 were also increased.Pathological examination showed that the normal structure of lung was destroyed badly after LPS injection.U0126 effectively inhibited the activation of ERK1/2,blocked LPS-induced NF-κB activation and HMGB-1 expression in lung tissue,reduced the lung water content[(4.59±0.51) vs (5.19±0.10),P＜0.05],the concentration of TNF-α,iNOS and IL-10[TNF-α,reached a peak at 3 h,(28±3) vs (70±10)(P＜0.05).iNOS,reached a peak at 12 h,(7 771±957) vs (10 679±1 641),P＜0.05.IL-10,reached a peak at 6 h,(59±10) vs (91±11),P＜0.05],and prevented LPS-induced lung injury.Conclusions ERK1/2 plays an important role in the development of LPS-induced ALI,and the activation of ERK1/2 may be one of the mechanisms of LPS-induced ALI.
