南方医科大学学报
南方醫科大學學報
남방의과대학학보
JOURNAL OF SOUTHERN MEDICAL UNIVERSITY
2009年
10期
1977-1981
,共5页
魏水生%廖新学%杨春涛%蔺际%杨战利%兰爱平%黄雪%王礼春%陈培熹%冯鉴强
魏水生%廖新學%楊春濤%藺際%楊戰利%蘭愛平%黃雪%王禮春%陳培熹%馮鑒彊
위수생%료신학%양춘도%린제%양전리%란애평%황설%왕례춘%진배희%풍감강
活性氧清除剂%心肌保护%化学性缺氧%线粒体膜电位%氧化应激
活性氧清除劑%心肌保護%化學性缺氧%線粒體膜電位%氧化應激
활성양청제제%심기보호%화학성결양%선립체막전위%양화응격
reactive oxygen species%cardioprotection%chemical hypoxia%mitochondrial membrane potential%oxidative stress
目的 探讨活性氧(ROS)清除剂N-乙酰半胱氨酸(NAC)能否保护H9c2心肌细胞对抗化学性缺氧引起的损伤.方法 应用化学性低氧模拟物氯化钴(CoCl_2)处理H9c2心肌细胞,建立化学性缺氧损伤心肌细胞的实验模型.在CoCl_2处理H9c2心肌细胞前60min把NAC加入培养基中,作为预处理.应用CCK-8比色法检测细胞存活率;双氯荧光素(DCFH-DA)染色荧光显微镜照相检测细胞内ROS水平;罗丹明123(Rh123)染色荧光显微镜照相检测线粒体膜电位(MMP);谷胱甘肽试剂盒检测GSSG/(GSSG+GSH)的比值.结果 600 μmol/L CoCl_2明显地降低细胞存活率.在CoCl_2处理H9c2心肌细胞前60 min,应用500～2000μmol/L NAC能剂量依赖性地抑制CoCl_2对心肌细胞的损伤作用,使细胞存活率显著升高.2000/μmol/LNAC能明显地对抗CoCl_2引起的氧化应激反应,使H9c2心肌细胞内GSSG/(GSSG+GSH)的比值及ROS水平明显降低,并明显地对抗CoCl_2对MMP的抑制作用.结论 NAC能显著地对抗化学性缺氧诱导的心肌细胞损伤,此心肌细胞保护作用与其降低GSSG/(GSSG+GSH)的比值及ROS水平,改善MMP等机制有关.
目的 探討活性氧(ROS)清除劑N-乙酰半胱氨痠(NAC)能否保護H9c2心肌細胞對抗化學性缺氧引起的損傷.方法 應用化學性低氧模擬物氯化鈷(CoCl_2)處理H9c2心肌細胞,建立化學性缺氧損傷心肌細胞的實驗模型.在CoCl_2處理H9c2心肌細胞前60min把NAC加入培養基中,作為預處理.應用CCK-8比色法檢測細胞存活率;雙氯熒光素(DCFH-DA)染色熒光顯微鏡照相檢測細胞內ROS水平;囉丹明123(Rh123)染色熒光顯微鏡照相檢測線粒體膜電位(MMP);穀胱甘肽試劑盒檢測GSSG/(GSSG+GSH)的比值.結果 600 μmol/L CoCl_2明顯地降低細胞存活率.在CoCl_2處理H9c2心肌細胞前60 min,應用500～2000μmol/L NAC能劑量依賴性地抑製CoCl_2對心肌細胞的損傷作用,使細胞存活率顯著升高.2000/μmol/LNAC能明顯地對抗CoCl_2引起的氧化應激反應,使H9c2心肌細胞內GSSG/(GSSG+GSH)的比值及ROS水平明顯降低,併明顯地對抗CoCl_2對MMP的抑製作用.結論 NAC能顯著地對抗化學性缺氧誘導的心肌細胞損傷,此心肌細胞保護作用與其降低GSSG/(GSSG+GSH)的比值及ROS水平,改善MMP等機製有關.
목적 탐토활성양(ROS)청제제N-을선반광안산(NAC)능부보호H9c2심기세포대항화학성결양인기적손상.방법 응용화학성저양모의물록화고(CoCl_2)처리H9c2심기세포,건립화학성결양손상심기세포적실험모형.재CoCl_2처리H9c2심기세포전60min파NAC가입배양기중,작위예처리.응용CCK-8비색법검측세포존활솔;쌍록형광소(DCFH-DA)염색형광현미경조상검측세포내ROS수평;라단명123(Rh123)염색형광현미경조상검측선립체막전위(MMP);곡광감태시제합검측GSSG/(GSSG+GSH)적비치.결과 600 μmol/L CoCl_2명현지강저세포존활솔.재CoCl_2처리H9c2심기세포전60 min,응용500～2000μmol/L NAC능제량의뢰성지억제CoCl_2대심기세포적손상작용,사세포존활솔현저승고.2000/μmol/LNAC능명현지대항CoCl_2인기적양화응격반응,사H9c2심기세포내GSSG/(GSSG+GSH)적비치급ROS수평명현강저,병명현지대항CoCl_2대MMP적억제작용.결론 NAC능현저지대항화학성결양유도적심기세포손상,차심기세포보호작용여기강저GSSG/(GSSG+GSH)적비치급ROS수평,개선MMP등궤제유관.
Objective To investigate the protective effect of reactive oxygen species (ROS) scavenger, N-acetyl-L-cysteine (NAC), against H9c2 cardiomyocytes from injuries induced by chemical hypoxia. Methods H9c2 cells were treated with cobalt chloride (CoCl_2), a chemical hypoxia-mimetic agent, to establish the chemical hypoxia-induced cardiomyocyte injury model. NAC was added into the cell medium 60 min prior to CoCl_2 exposure. The cell viability was evaluated using cell counter kit (CCK-8), and the intercellular ROS level was measured by 2', 7'- dichlorfluorescein-diacetate (DCFH-DA) staining and photofluorography. Mitochondrial membrane potential (MMP) of the cells was observed by Rhodamine 123 (Rh123) staining and photofluorography, and the ratio of GSSG/ (GSSG+GSH) was calculated according to detection results of the GSSG kit.Results Exposure of H9c2 cardiomyocytes to 600 μmol/L CoCl_2 for 36 h resulted in significantly reduced cell viability. Pretreatment with NAC at the concentrations ranging from 500 to 2000 μmol/L 60 min before CoCl_2 exposure dose-dependently inhibited CoCl_2-induced H9c2 cell injuries, and obviously increased the cell viability. NAC at 2000 μmol/L obviously inhibited the oxidative stress induced by CoCl_2, decreased the ratio of GSSG/(GSSG+GSH), increased ROS level, and antagonized CoCl_2-induced inhibition on MMP. Conclusions NAC offers obvious protective effect on H9c2 cardiomyocytes against injuries induced by chemical hypoxia by decreasing in the ratio of GSSG/ (GSSG+GSH) and ROS level and ameliorating MMP.
