中华实用儿科临床杂志
中華實用兒科臨床雜誌
중화실용인과림상잡지
Journal of Applied Clinical Pediatrics
2014年
18期
1372-1376
,共5页
陈扬%陆铸今%杨毅%陆国平
陳颺%陸鑄今%楊毅%陸國平
진양%륙주금%양의%륙국평
急性肺损伤%纤溶酶原激活物抑制剂-1%一氧化氮%一氧化氮合酶
急性肺損傷%纖溶酶原激活物抑製劑-1%一氧化氮%一氧化氮閤酶
급성폐손상%섬용매원격활물억제제-1%일양화담%일양화담합매
Acute Iung injury%Plasminogen activator inhibitor-1%Nitric oxide%Nitric oxide synthase
目的 探讨早期吸入一氧化氮(NO)对急性肺损伤大鼠纤溶酶原激活物抑制剂-1(PAI-1)mRNA和蛋白表达的影响及其意义;观察吸入NO后急性肺损伤大鼠诱生性NO合酶(iNOS)和内源性NO的变化及其与PAI-1表达的关系.方法 采用内毒素(LPS)二次打击方法建立4～5周SD大鼠急性肺损伤模型.对照组和LPS组分别随机给予吸入空气(A)、20×10-6 NO,干预24 h.应用荧光实时定量PCR方法测定大鼠肺组织PAI-1 mRNA水平,免疫组织化学测定2组大鼠肺组织PAI-1蛋白的表达水平,并测定肺组织iNOS活性和NO水平;同时行肺组织病理评分和纤维素染色.结果 造模后气体干预24 h时肺组织PAI-1 mRNA和蛋白表达在NO干预的LPS-NO组较LPS-A组显著降低(4.94 ±0.52比5.56±0.27;1.31 ±0.40比1.69 ±0.16,P均＜0.05).同时NO干预后LPS-NO组iNOS活性和肺组织NO水平均显著低于LPS-A组[(0.84±0.36)U/mg prot比(2.30±0.25) U/mg prot;(1.90±0.84) μmol/g prot比(3.38±0.73) μmol/g prot,P均＜0.05).iNOS活性与PAI-1 mRNA和蛋白表达呈正相关(r=0.481,P=0.005;r =0.667,P=0.000);肺组织NO水平与PAI-1 mRNA和蛋白表达呈正相关(r=0.532,P=0.002;r =0.784,P=0.000).肺病理评分在干预24 h时,LPS-NO组较LPS-A组下降,差异有统计学意义(4.28±0.94比6.12±1.51,P＜0.05).光镜下LPS-NO组大鼠肺纤维素沉积较吸入空气的大鼠有所减少.结论 早期吸入20×10-6 NO可抑制急性肺损伤大鼠PAI-1的高表达,缓解纤溶失衡,减少纤维蛋白沉积,减轻肺损伤;吸入NO可减少急性肺损伤时肺组织iNOS活性和NO产量,此作用与PAI-1表达下调密切相关,因此可将研究内源性NO系统调节PAI-1表达的信号通路作为下一步的研究方向,为设计新疗法提供思路.
目的 探討早期吸入一氧化氮(NO)對急性肺損傷大鼠纖溶酶原激活物抑製劑-1(PAI-1)mRNA和蛋白錶達的影響及其意義;觀察吸入NO後急性肺損傷大鼠誘生性NO閤酶(iNOS)和內源性NO的變化及其與PAI-1錶達的關繫.方法 採用內毒素(LPS)二次打擊方法建立4～5週SD大鼠急性肺損傷模型.對照組和LPS組分彆隨機給予吸入空氣(A)、20×10-6 NO,榦預24 h.應用熒光實時定量PCR方法測定大鼠肺組織PAI-1 mRNA水平,免疫組織化學測定2組大鼠肺組織PAI-1蛋白的錶達水平,併測定肺組織iNOS活性和NO水平;同時行肺組織病理評分和纖維素染色.結果 造模後氣體榦預24 h時肺組織PAI-1 mRNA和蛋白錶達在NO榦預的LPS-NO組較LPS-A組顯著降低(4.94 ±0.52比5.56±0.27;1.31 ±0.40比1.69 ±0.16,P均＜0.05).同時NO榦預後LPS-NO組iNOS活性和肺組織NO水平均顯著低于LPS-A組[(0.84±0.36)U/mg prot比(2.30±0.25) U/mg prot;(1.90±0.84) μmol/g prot比(3.38±0.73) μmol/g prot,P均＜0.05).iNOS活性與PAI-1 mRNA和蛋白錶達呈正相關(r=0.481,P=0.005;r =0.667,P=0.000);肺組織NO水平與PAI-1 mRNA和蛋白錶達呈正相關(r=0.532,P=0.002;r =0.784,P=0.000).肺病理評分在榦預24 h時,LPS-NO組較LPS-A組下降,差異有統計學意義(4.28±0.94比6.12±1.51,P＜0.05).光鏡下LPS-NO組大鼠肺纖維素沉積較吸入空氣的大鼠有所減少.結論 早期吸入20×10-6 NO可抑製急性肺損傷大鼠PAI-1的高錶達,緩解纖溶失衡,減少纖維蛋白沉積,減輕肺損傷;吸入NO可減少急性肺損傷時肺組織iNOS活性和NO產量,此作用與PAI-1錶達下調密切相關,因此可將研究內源性NO繫統調節PAI-1錶達的信號通路作為下一步的研究方嚮,為設計新療法提供思路.
목적 탐토조기흡입일양화담(NO)대급성폐손상대서섬용매원격활물억제제-1(PAI-1)mRNA화단백표체적영향급기의의;관찰흡입NO후급성폐손상대서유생성NO합매(iNOS)화내원성NO적변화급기여PAI-1표체적관계.방법 채용내독소(LPS)이차타격방법건립4～5주SD대서급성폐손상모형.대조조화LPS조분별수궤급여흡입공기(A)、20×10-6 NO,간예24 h.응용형광실시정량PCR방법측정대서폐조직PAI-1 mRNA수평,면역조직화학측정2조대서폐조직PAI-1단백적표체수평,병측정폐조직iNOS활성화NO수평;동시행폐조직병리평분화섬유소염색.결과 조모후기체간예24 h시폐조직PAI-1 mRNA화단백표체재NO간예적LPS-NO조교LPS-A조현저강저(4.94 ±0.52비5.56±0.27;1.31 ±0.40비1.69 ±0.16,P균＜0.05).동시NO간예후LPS-NO조iNOS활성화폐조직NO수평균현저저우LPS-A조[(0.84±0.36)U/mg prot비(2.30±0.25) U/mg prot;(1.90±0.84) μmol/g prot비(3.38±0.73) μmol/g prot,P균＜0.05).iNOS활성여PAI-1 mRNA화단백표체정정상관(r=0.481,P=0.005;r =0.667,P=0.000);폐조직NO수평여PAI-1 mRNA화단백표체정정상관(r=0.532,P=0.002;r =0.784,P=0.000).폐병리평분재간예24 h시,LPS-NO조교LPS-A조하강,차이유통계학의의(4.28±0.94비6.12±1.51,P＜0.05).광경하LPS-NO조대서폐섬유소침적교흡입공기적대서유소감소.결론 조기흡입20×10-6 NO가억제급성폐손상대서PAI-1적고표체,완해섬용실형,감소섬유단백침적,감경폐손상;흡입NO가감소급성폐손상시폐조직iNOS활성화NO산량,차작용여PAI-1표체하조밀절상관,인차가장연구내원성NO계통조절PAI-1표체적신호통로작위하일보적연구방향,위설계신요법제공사로.
Objective To explore the effects of inhaled nitric oxide(NO) on expression of plasminogen activator inhibitor-1 (PAI-1) mRNA and protein in the early-stage of experimental acute lung injury (ALI) in a rat model.And to investigate the relationship between endogenous NO system including inducible nitric oxide synthase (iNOS)and intrapulmonary NO production and expressions of PAI-1 in ALI.Methods In the study,endotoxemia followed by the second attack due to intratracheal injection of lipopolysaccharide (LPS) in rats caused ALI.Male SD rats aged 4-5 weeks (clean conventional rats,180-200 g) were randomly assigned to 2 groups:saline control (C) group,LPS-treated (LPS) group,and the 2 groups were randomly allocated to subgroups exposed to air (A) or 20 × 10-6 NO.They were sacrificed for 24 h.Expressions of PAI-1 mRNA of the lung tissue were evaluated by real-time polymerise chain reaction; PAI-1 proteins were determined by immunohistochemistry.NO production in the lung tissues and pulmonary iNOS activity were measured.Meanwhile,histopathological lung injury scores were evaluated and modified martius acid fuchsin brilliant blue(MSB) stains was performed to evaluate fibrin of the lung tissues.Results At 24 h time point with intervention of iNO,PAI-1 mRNA and protein levels in LPS-NO subgroup were decreased compared with those in LPS-A subgroup (4.94 ± 0.52 vs 5.56 ± 0.27 ; 1.31 ± 0.40 vs 1.69 ± 0.16,all P ＜ 0.05).Meanwhile,iNOS activity and NO productions in LPS-NO subgroup were lower than those of LPS-A subgroup [(0.84 ± 0.36) U/mg prot vs (2.30 ± 0.25) U/mg prot ; (1.90 ± 0.84) μmol/g prot vs (3.38 ± 0.73) μmol/g prot,all P ＜ 0.05].iNOS activity had significant correlation with expression of PAI-1 mRNA and protein in lung tissue (r =0.481,P =0.005 ; r =0.667,P =0.000) ; NO production had significant correlation with expression of PAI-1 mRNA and protein in lung tissue(r =0.532,P =0.002; r =0.784,P =0.000).At 24 h time point,the histopathologic lung injury scores in LPS-NO subgroup were decreased in contrast to LPS-A subgroup (4.28 ±0.94 vs 6.12 ± 1.51,P ＜ 0.05).Fibrin deposition evaluated by modified MSB stains in LPS subgroups was found in alveolar space,lumen of blood vessel and mesenchymal ;LPS subgroup with NO appeared a decreasing trend in contrast to LPS subgroup with air.Conclusions Inhaled nitric oxide of 20 × 10 6 can suppress elevated expression of PAI-1 in ALI induced by endotoxin.This inhaled NO can improve the imba-lance of plasminogen activation system and alleviate lung injury.Meanwhile,inhaled NO down-regulates intrapulmonary iNOS activity as well as endogenous NO productions in rats with 2 hits of LPS induced ALI.These changes also have a close correlation with down-regulation of PAI-1 mRNA and protein.Thus,regulation of endogenous NO system on the expression of PAI-1 will be the future direction of new therapies for ALI.
