摘要
目的观察硫化氢(hydrogen sulfide,H2S)供体——硫氢化钠(sodium hydrosulfide,NaHS)后处理对培养的新生大鼠心肌细胞缺氧/复氧损伤中线粒体渗透性转换(mitochon-drial permeable transition,MPT)影响。方法将原代培养大鼠乳鼠心肌细胞随机分为缺氧/复氧组(H/R)和硫氢化钠后处理组(H/R+NaHS)。H/R组为缺氧2h后复氧2h。H/R+NaHS组为缺氧2h后,给予1×10-6mol·L-1NaHS后处理0.5h,再复氧1.5h。两组分别于缺氧前、缺氧2h、复氧0.5h、1和2h,应用噻唑蓝法检测细胞活性,荧光指示剂孵育激光共聚焦显微镜下检测线粒体渗透性转换孔(MPTP)的开放程度和线粒体膜电位(Δψm)的变化。结果缺氧2h内细胞活力明显下降,MPTP开放,Δψm下降。在复氧初期两组细胞损伤会进一步加剧,但H/R+NaHS组在NaHS后处理后,细胞活性(45%±3.7%)下降幅度明显小于H/R组(36.5%±1.8%,P<0.05);MPTP荧光强度(52%±5.7%)和Δψm荧光强度(47.2%±5.1%)也明显高于H/R组(40%±5.4%,33.4%±5.0%,P<0.05)。到复氧2h时,两组细胞损伤均不再加重,但H/R+NaHS组各项指标仍然明显好于H/R组(P<0.05)。结论硫氢化钠后处理可以有效调控缺氧/复氧心肌细胞线粒体渗透性转换,阻止MPTP开放,抑制Δψm下降,从而减轻缺氧/复氧造成的心肌细胞损伤。
Aim To explore the effect of NariS （ the donator of H2S ） postconditioning on the mitochondrial permeable transition （MPT） in cultured neonatal rat myocardial cells exposed to hypoxia/reoxygenation. Methods Primary cultured myocardial cells from neonatal rats were randomly divided into hypoxia/reoxygenation group （H/R） and Naris postconditioning group （H/R ＋ NaHS ）. The cardiomyocytes in H/R group were made hypoxia for 2 hours followed by 2 hours reoxygenation, After 2 hours hypoxia cardiomyocytes in H/R ＋ NaHS group were treated with 1 × 10^-6 mol· L^-1 Naris postconditioning for 30 minutes, then followed by 1.5 hours reoxygenation. Some items were detected at pre-hypoxia,hypoxia 2 h and reoxygenation 0.5 h, 1 h and 2 h in both groups. The cell viability was detected by MTT method. The opening of mitochondrial permeable transition pore （MPTP） and the △ψm change were measured by fluorescent dyes with confocal microscopy. Results During 2 hours hypoxia, cell viability and △ψm descended significantly, and MPTP began to open. In the early reoxygenation, the injury of cardiomyocytes aggravated. But after NariS postconditioning, cell viability （ 45% ± 3.7% ） was higher than that of H/R group （36.5% ± 1.8% ,P 〈 0. 05 ）, and fluorescent intensity of Calcein AM and TMRE （52% ±5.7%,47.2% ± 5.1%） was also higher than that of H/R group （40% ±5.4% ,33.4% ± 5.0% , P 〈 0. 05 ）. However these changes in two groups inclined to stabilize at the end of 2 h reoxygenation but all results in those of H/R group were also better than those of H/R ＋ Naris group. Conclusions NariS postconditioning might effectively regulate MPT in cultured rat myocardial cells during hypoxia/ reoxygenation, which could prevent the opening of MPTP,inhibit the decline of △ψm and thus relieve cardiac injury.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2008年第12期1610-1614,共5页
Chinese Pharmacological Bulletin
基金
国家自然科学基金资助项目(No30571782)
关键词
硫化氢
缺氧/复氧
后处理
线粒体渗透性转换
hydrogen sulfide
hypoxia/reoxygenation
postconditioning
mitochondrial permeable transition
