摘要
目的 探讨硫化氢后处理对大鼠心肌缺血再灌注时左心室收缩功能的影响.方法 实验一成年雄性SD大鼠,体重200～250 g,采用Langendorff装置建立离体心脏灌注模型.取离体灌注模型制备成功的心脏40个,随机分为5组（n=8）：对照组（C组）、缺血再灌注组（IR组）和硫氢化钠1、10、100 μmol/L后处理组（SP1组、SP10组、SP100组）.平衡灌注20 min后,C组继续灌注K-H液100 min;IR组灌注ST.Thomas停跳液10 ml/kg后停灌40 min,恢复K-H液灌注60 min;SP1组、SP10组、SP100组全心缺血40 min,于再灌注前灌注含1、10、100 μmol/L硫氢化钠的K-H液2 min,然后恢复K-H液灌注60 min.于平衡灌注末和再灌注60 min时,记录左室发展压（LVDP）、左心室发展压最大上升速率（＋dp/dtmax）和左心室发展压最大下降速率（-dp/dtmax）.实验二成年雄性SD大鼠,体重200～250 g,分离心肌细胞,加入培养皿中,放入95%O2-5%CO2培养箱中培养4 h.取64皿细胞,随机分为4组（n=16）：对照组（C组）、缺氧复氧组（HR组）、硫化氢后处理组（SP组）和缺氧后处理组（HP组）.C组继续于95%O2-5%CO2培养箱中培养2 h;HR组于95%N3-5%CO2培养箱中缺氧1 h,然后于95%O2-5%CO2培养箱中复氧1 h;SP组于95%N2-5%CO2培养箱中缺氧1 h,加入硫氢化钠10μmol/L孵育2 min,然后于95%O2-5%CO2培养箱中复氧1 h;HP组于95%N2-5%CO2培养箱中缺氧1 h,然后复氧3 min,缺氧3 min,重复3次,再于95%O2-5%CO2培养箱中复氧1 h.测定线粒体膜电位和F-肌动蛋白的表达.结果 实验一与C组比较,再灌注60 min时IR组LVDP和±dp/dp/dtmax降低（P＜0.05）,SP1组、SP10组和SP100组LVDP和±dp/dtmax差异无统计学意义（P＞0.05）;与IR组比较,SP1组、SP10组和SP100组LVDP和±dp/dtmax升高（P＜0.05）;SP1组、SP10组和SP100组间LVDP和±dp/dtmax比较差异无统计学意义（P＞0.05）.实验二与C组比较,HR组和HP组线粒体膜电位降低,HR组、SP组和HP组F-肌动蛋白表达上调（P＜0.05）;与HR组比较,SP组和HP组线粒体膜电位升高,F-肌动蛋白表达上调（P＜0.05）;SP组和HP组间线粒体膜电位和F-肌动蛋白表达比较差异无统计学意义（P＞0.05）.结论硫化氢后处理可改善大鼠心肌缺血再灌注时左心室收缩功能,与其稳定心肌细胞线粒体膜电位和促进F-肌动蛋白聚集有关.
Objective To investigate the effect of hydrogen sulfide postconditioning on the systolic function of left ventricle during myocardial ischemia-reperfusion （IR） in rats. Methods Part Ⅰ Adult male SD rats weighing 200-250 g were anesthetized with pentobarbital 40 mg/kg and heparin 250 U/kg. Their hearts were excised and perfused in a Langendorff apparatus with K-H solution saturated with 95% O2-5% CO2 at 37 ℃. Forty isolated rat hearts were randomly divided into 5 groups （ n = 8 each） after 20 min of equilibration： control group （group C）; IR group; sodium hydrosulfide 1,10, 100 μmol/L postconditioning group （group SP1, SP10, SP100 ）.In group Cthe hearts were perfused continuously for another 100 min. In group IR, the hearts were reperfused for 60 min after 40 min ischemia induced by 10 ml/kg ST. Thomas solution. In group SP1 , SP10 and SP100 the hearts were perfused with K-H solution containing sodium hydrosulfide 1, 10, 100 μmol/L for 2 min before reperfusion.LVDP and ± dp/dtmax were recorded at the end of equilibration and reperfusion. Part Ⅱ Cardiomyocytes were isolated from the male SD rats （weighing 200-250 g） and then cultured in CO2 incubator for 4 h. Sixty-four dishes of cultured myocytes were randomly divided into 4 groups（ n = 16 each）： control group （group C）, hypoxia/reoxygenation group （group HR）, hydrogen sulfide postconditioning group （group SP） and hypoxia postconditioning group （group HP）. Group C were cultured continuously for 2 h. Group HR, SP and HP were exposed to 1 h hypoxia （95%N2-5%CO2 ） followed by 1 h reoxygenation. In group SP 10 μmol/L sodium hydrosulfide was added and the myocytes were then incubated for 2 min before reoxygenation. In group HP the cultured myocytes were expased to 3 min reoxygenation followed by 3 min hypoxia for 3 times before the 1 h reoxygenation. Mitochondrial membrane potential and F-actin expression were determined. Results Part Ⅰ Compared with group C, LVDP and ± dp/dtmax were significantly decreased at the end of reperfusion in group IR （P 〈 0.05）, while no significant difference was found in group SP1 , SP10 and SP100（P 〉0.05）. Compared with group IR, LVDP and ± dp/dtmax were significantly increased in group SP （ P 〈 0.05）. There was no significant difference in LVDP and ± dp/dtmax among group SP1, SP10 and SP100（P 〉0.05）. Part H Compared with group C, the mitochondrial membrane potential was significantly decreased in group HR and HP, and the expression of F-actin was significantly up-regulated in group HR, SP and HP （ P 〈 0.05）. Compared with group HR, the mitochondrial membrane potential was significantly increased and the expression of F-actin up-regulated in group SP and HP （ P 〈 0.05 ）. There were no significant difference in the mitochondrial membrane potential and expression of F-actin between group SP and HP （ P 〉0.05）.Conclusion Hydrogen sulfide postconditioning can improve left ventricular systolic function during IR in rats by stabilizing mitochondrial membrane potential and promoting aggregation of F-actin.
出处
《中华麻醉学杂志》
CAS
CSCD
北大核心
2010年第10期1175-1178,共4页
Chinese Journal of Anesthesiology
基金
卫生部卫生公益性行业科研专项项目（200802173）
关键词
硫化氢
心肌再灌注损伤
心脏功能试验
Hydrogen sulfide
Myocardial reperfusion injury
Heart function tests
