南方医科大学学报
南方醫科大學學報
남방의과대학학보
JOURNAL OF SOUTHERN MEDICAL UNIVERSITY
2011年
8期
1289-1294
,共6页
不对称二甲基精氨酸%二甲基精氨酸一二甲胺水解酶%急性肺损伤%鼠脑缺血再灌注
不對稱二甲基精氨痠%二甲基精氨痠一二甲胺水解酶%急性肺損傷%鼠腦缺血再灌註
불대칭이갑기정안산%이갑기정안산일이갑알수해매%급성폐손상%서뇌결혈재관주
asymmetric dimethylarginine%dimethylarginine dimethylaminohydrolase%acute lung injury%cerebral ischemia/reperfusion injury
目的 研究非对称二甲基精氨酸(ADMA)在大鼠脑缺血再灌注(I/R)诱导的急性肺损伤发病过程中的作用.方法 成年雄性SD大鼠随机分为假手术组(S)、模型组(I/R)、ADMA处理组(ADMA+I/R)和DDAH处理组(DDAH+I/R).通过大鼠脑缺血2h后恢复血流灌注诱导急性肺损伤.在恢复血流灌注24 h后,采用比色法检测各组大鼠肺组织一氧化氮合酶(NOS)活性和NO含量.采用RT-PCR和Western blotting分别检测肺组织蛋白激酶(PKC)和肌球蛋白轻链激酶(MLCK)mRNA和蛋白表达水平；ELISA法检测支气管肺泡灌洗液及入肺血和出肺血血浆中的ADMA水平.结果 脑I/R损伤大鼠支气管肺泡灌洗液和入肺血血浆中ADMA水平明显升高,肺组织中NO含量和NOS活性明显降低(P<0.05),同时MLCK和PKC mRNA和蛋白表达明显上调(P<0.05).预先给予外源性DDAH后,脑缺血/再灌注损伤大鼠支气管肺泡灌洗液和人肺血血浆中ADMA水平明显降低,肺组织NO含量和NOS活性明显升高,MLCK和PKC mRNA和蛋白表达明显下调（P<0.05）.结论 ADMA通过上调肺组织MLCK和PKC的表达参与了脑I/R损伤后急性肺损伤的发病过程.ADMA可能是脑缺血／再灌注损伤诱导急性肺损伤的—个新的生物标志物和治疗靶点.
目的 研究非對稱二甲基精氨痠(ADMA)在大鼠腦缺血再灌註(I/R)誘導的急性肺損傷髮病過程中的作用.方法 成年雄性SD大鼠隨機分為假手術組(S)、模型組(I/R)、ADMA處理組(ADMA+I/R)和DDAH處理組(DDAH+I/R).通過大鼠腦缺血2h後恢複血流灌註誘導急性肺損傷.在恢複血流灌註24 h後,採用比色法檢測各組大鼠肺組織一氧化氮閤酶(NOS)活性和NO含量.採用RT-PCR和Western blotting分彆檢測肺組織蛋白激酶(PKC)和肌毬蛋白輕鏈激酶(MLCK)mRNA和蛋白錶達水平；ELISA法檢測支氣管肺泡灌洗液及入肺血和齣肺血血漿中的ADMA水平.結果 腦I/R損傷大鼠支氣管肺泡灌洗液和入肺血血漿中ADMA水平明顯升高,肺組織中NO含量和NOS活性明顯降低(P<0.05),同時MLCK和PKC mRNA和蛋白錶達明顯上調(P<0.05).預先給予外源性DDAH後,腦缺血/再灌註損傷大鼠支氣管肺泡灌洗液和人肺血血漿中ADMA水平明顯降低,肺組織NO含量和NOS活性明顯升高,MLCK和PKC mRNA和蛋白錶達明顯下調（P<0.05）.結論 ADMA通過上調肺組織MLCK和PKC的錶達參與瞭腦I/R損傷後急性肺損傷的髮病過程.ADMA可能是腦缺血／再灌註損傷誘導急性肺損傷的—箇新的生物標誌物和治療靶點.
목적 연구비대칭이갑기정안산(ADMA)재대서뇌결혈재관주(I/R)유도적급성폐손상발병과정중적작용.방법 성년웅성SD대서수궤분위가수술조(S)、모형조(I/R)、ADMA처리조(ADMA+I/R)화DDAH처리조(DDAH+I/R).통과대서뇌결혈2h후회복혈류관주유도급성폐손상.재회복혈류관주24 h후,채용비색법검측각조대서폐조직일양화담합매(NOS)활성화NO함량.채용RT-PCR화Western blotting분별검측폐조직단백격매(PKC)화기구단백경련격매(MLCK)mRNA화단백표체수평；ELISA법검측지기관폐포관세액급입폐혈화출폐혈혈장중적ADMA수평.결과 뇌I/R손상대서지기관폐포관세액화입폐혈혈장중ADMA수평명현승고,폐조직중NO함량화NOS활성명현강저(P<0.05),동시MLCK화PKC mRNA화단백표체명현상조(P<0.05).예선급여외원성DDAH후,뇌결혈/재관주손상대서지기관폐포관세액화인폐혈혈장중ADMA수평명현강저,폐조직NO함량화NOS활성명현승고,MLCK화PKC mRNA화단백표체명현하조（P<0.05）.결론 ADMA통과상조폐조직MLCK화PKC적표체삼여료뇌I/R손상후급성폐손상적발병과정.ADMA가능시뇌결혈／재관주손상유도급성폐손상적—개신적생물표지물화치료파점.
Objective To determine the role of asymmetric dimethylarginine (ADMA) in acute lung injury induced by cerebral ischemia/reperfusion (I/R) injury in rats.Methods Adult male SD rats were randomly divided into 4 groups,namely the sham-operated group (S),cerebral I/R model group,ADMA + I/R group,and dimethylarginine dimethylaminohydrolase (DDAH)+I/R group.In the latter 3 groups,acute lung injury was induced by left middle cerebral artery occlusion for 120 min.After a 24-h reperfusion,the rats were sacrificed and the activities of nitric oxide synthase (NOS) and contents of nitric oxide (NO) were measured using reductase and colorimetric assay.The mRNA and protein expressions of protein kinase C (PKC)and myosin light chain kinase (MLCK) in the lung tissues were detected with RT-PCR and Western blotting,respectively.The contents of ADMA in the bronchoalveolar lavage fluid (BALF) and blood flowing into and out of the lungs were measured by ELISA.Results Cerebral I/R injury caused significantly elevated ADMA levels in the BALF and blood flowing into the lungs,and obviously lowered the NO concentration and NOS activity in the lung tissues (P＜0.05).Following cerebral I/R injury,MLCK and PKC mRNA and protein expressions were significantly upregualted in the lung tissues (P＜O.05).Exogenous DDAH obviously decreased the levels of ADMA in the BALF and blood flowing into the lungs,increased NO concentration and NOS activity,and down-reguahed MLCK and PKC mRNA and protein expressions in lung tissues of rats with cerebral I/R injury (P＜0.05).Conclusion ADMA contributes to the development of acute lung injury following cerebral I/R injury in rats by upregulating MLCK and PKC expression.ADMA may serve as a novel therapeutic biomarker and a potential therapeutic target for acute lung injury induced by cerebral I/R injury.
