摘要
目的研究吗啡预处理对心肌的保护作用以及格列苯脲对其的影响。方法采用新生大鼠的原代心肌培养细胞作为研究对象,将培养心肌细胞随机分配到4组:I组,空白对照组,共有10瓶细胞,该组细胞没有进行缺氧再复氧;II组,缺氧再复氧组,共有10瓶细胞,该组细胞进行缺氧条件下培养6h再有氧条件下培养18h;III组,吗啡预处理组,与II组细胞基本相同,唯一区别是在缺氧再复氧之前,向细胞培养瓶内加入吗啡,使细胞培养瓶内培养基中含有1μmol/L浓度的吗啡,15min后,再进行缺氧再复氧。IV组,格列苯脲组,首先在缺氧再复氧之前,向细胞培养瓶内加入格列苯脲,使细胞培养瓶内培养基中含有100μmol/L浓度的格列苯脲,15min后,再向细胞培养瓶内加入吗啡,使细胞培养瓶内培养基中含有1μmol/L浓度的吗啡,15min后,再进行缺氧再复氧。最后所有细胞进行电镜检查,并且通过PI染色的流式细胞术来检测细胞凋亡情况。结果缺氧再复氧组细胞在经历6h的缺氧培养,再18h的有氧培养后,细胞大部分出现凋亡,与空白对照组细胞相比,有明显的升高(P<0.05),而吗啡预处理组细胞凋亡指数与缺氧再复氧组相比有明显的降低(P<0.05)。但是格列苯脲组细胞的凋亡指数与吗啡组相比有明显升高(P<0.05),与缺氧再复氧组相比没有明显差别。根据电镜结果,格列苯脲组细胞与缺氧再复氧组细胞线粒体和内质网有明显的空泡变性,细胞核浓缩,而吗啡预处理组细胞形态基本正常。结论吗啡预处理可以明显抑制缺氧再复氧损伤所引起的细胞凋亡,格列苯脲可以拮抗吗啡的心肌保护作用。
[Objective] To study the effect of morphine on apoptosis induced by hypoxia-reoxygenation in murine cultured heart cells and glibenclamide effect on morphine preconditioning. [Methods] Cultured neonatal rat cardiac myocytes were prepared and randomly assigned into four groups： Group Ⅰ , the control group; Group Ⅱ, the hypoxia-reoxygenation group; Group Ⅲ, the morphine-preconditioning group; and Group Ⅳ, morphine plus glibenclamide group. The morphology of these cells were observed by transmission electron microscopy and the apoptosis index （AI） of these cells were assessed by fluorescence labelling with Propidium Iodide （PI） and analysed by flow cytometry system. [Results] The apoptosis index（AI） in Group Ⅱ was significantly higher than that in Group Ⅰ （P 〈0.05）. While the morphine-preconditioning cells showed a descending apoptosis index compared with Group Ⅱ （P 〈0.05）, but the cells in Group Ⅳ showed the same AI level as Group Ⅱ. [Conclusion] Morphine has protective effect on apoptosis induced by hypoxia-reoxygenation in murine cultured heart cells and glibenclamide can abolish the cardioprotection of morphine.
出处
《中国现代医学杂志》
CAS
CSCD
北大核心
2008年第5期547-550,共4页
China Journal of Modern Medicine
基金
中南大学湘雅三医院博士化基金资助课题(No:0411)
关键词
吗啡
格列苯脲
凋亡
心肌
morphine
glibenclamide
apoptosis
myocyte
