中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (11): 1948-1953.doi: 10.3969/j.issn.1673-8225.2012.11.012

• 组织构建细胞学实验 cytology experiments in tissue construction • 上一篇    下一篇

阿魏酸钠预处理保护成年大鼠心肌细胞缺氧复氧的损伤★

曾  梁,刘季春   

  1. 南昌大学第一附属医院,江西省南昌市  330006
  • 收稿日期:2011-10-29 修回日期:2012-01-19 出版日期:2012-03-11
  • 通讯作者: 刘季春,博士,主任医师,南昌大学第一附属医院,江西省南昌市 330006 liujichun999@yahoo.com.cn
  • 作者简介:曾梁★,男,1980年生,江西省泰和县人,2006年南昌大学医学院毕业,硕士,主治医师,主要从事心脏相关基础与临床研究(心肌保护等)。zengliang0811@hotmail.com

Protective effect of sodium ferulate preconditioning on anoxia-reoxygenation injury of myocardial cells in adult rats

Zeng Liang, Liu Ji-chun   

  1. First Affiliated Hospital of Nanchang University, Nanchang  330006, Jiangxi Province, China
  • Received:2011-10-29 Revised:2012-01-19 Online:2012-03-11
  • Contact: author: Liu Ji-chun, Doctor, Chief physician, First Affiliated Hospital of Nanchang University, Nanchang 330006, Jiangxi Province, China liujichun999@yahoo.com.cn
  • About author:Zeng Liang★, Master, Attending physician, First Affiliated Hospital of Nanchang University, Nanchang 330006, Jiangxi Province, China zengliang0811@hotmail.com

摘要:

背景:阿魏酸钠预处理能对心肌细胞产生保护作用并已在大鼠体外心脏和乳鼠心肌细胞水平得以证实,尚缺乏在成年大鼠心肌细胞水平上的研究。
目的:探讨阿魏酸钠预处理对成年大鼠缺氧复氧心肌细胞的保护作用及其K+ATP通道机制。
方法:用Langendorff系统灌流心脏,以胶原酶消化法分离纯化成年大鼠心肌细胞并给予模拟缺氧复氧液以及干预因素阿魏酸钠、K+ATP通道阻断剂格列本脲处理,随机分为6组:正常对照组、缺氧复氧组、缺氧预适应+缺氧复氧组、阿魏酸钠+缺氧复氧组、格列本脲+缺氧预适应+缺氧复氧组、格列本脲+阿魏酸钠+缺氧复氧组。
结果与结论:①心肌细胞存活率:缺氧复氧组与对照组比较,细胞存活率明显降低(P < 0.01);缺氧预适应+缺氧复氧组、阿魏酸钠+缺氧复氧组与缺氧复氧组比较,细胞存活率明显增高(P < 0.01);格列本脲+缺氧预适应+缺氧复氧组、格列本脲+阿魏酸钠+缺氧复氧组与缺氧复氧组相比,差异无显著性意义,但明显高于缺氧预适应+缺氧复氧组(P < 0.01)。②乳酸脱氢酶活性:各组间乳酸脱氢酶活性比较结果与心肌细胞存活率比较结果吻合。③透射电镜观察:缺氧复氧组心肌细胞线粒体明显肿胀,嵴消失或变形,细胞膜结构破坏,有核边聚现象;缺氧预适应+缺氧复氧组、阿魏酸钠+缺氧复氧组心肌细胞超微结构改变轻微,线粒体排列规则、大小均匀,嵴及内外膜清晰完整,核膜完整,较缺氧复氧组损伤明显减轻;格列本脲+缺氧预适应+缺氧复氧组、格列本脲+阿魏酸钠+缺氧复氧组心肌细胞超微结构改变与缺氧复氧组相似。结果可见阿魏酸钠预处理对成年大鼠心肌细胞具有药理性心肌缺血预适应保护作用且该作用可能与K+ATP通道有关。
关键词:阿魏酸钠;成年大鼠;心肌细胞;缺氧复氧;心肌保护
doi:10.3969/j.issn.1673-8225.2012.11.012

关键词: 阿魏酸钠, 成年大鼠, 心肌细胞, 缺氧复氧, 心肌保护

Abstract:

BACKGROUND: Sodium ferulate preconditioning can protect myocardial cells, which has been proved in the experiments of isolated rat heart and neonatal rat myocardial cells, but there are few experimental studies regarding adult rat myocardial cells.
OBJECTIVE: To explore the protective effect of sodium ferulate preconditioning on myocardial cells following anoxia-reoxygenation (A/R) in adult rats as well as its mechanism underlying adenosine triphosphate-sensitive potassium (K+ATP) channel .
METHODS: Rat heart was perfused with Langendorff system. The purified adult rat myocardial cells were isolated by collagenase digestion method and then treated with simulated A/R solution, sodium ferulate (SF) as well as glibenclamide (Glib), a K+ATP channel blocker. Then, the myocardial cells were randomly divided into six groups: control group, A/R group, anoxia preconditioning (APC)+A/R group, SF+A/R group, Glib+APC+A/R group and Glib+SF+A/R group.
RESULTS AND CONCLUSION: ①Myocardial cell viability: compared with the control group, the cell viability in the A/R group was significantly decreased (P < 0.01). Compared with the A/R group, the cell viability was significantly increased in the APC+A/R group and SF+A/R group (P < 0.01). The cell viability in the Glib+APC+A/R and Glib+SF+A/R groups had no difference from that of the A/R group, but significantly higher than that of the APC+A/R group (P < 0.01). ②Lactate dehydrogenase activity: the comparison results of lactate dehydrogenase activity were consistent with those of myocardial cell viability among these groups. ③Ultrastructure observation under a transmission electron microscope: the myocardial cells subjected to A/R injury were characterized by mitochondrial swelling, disappearance or deformation of mitochondrial cristae, disruption of nuclear membrane, and nuclear condensation. While the ultrastructure of the myocardial cells in the APC+A/R group and SF+A/R group were changed slightly, exhibiting regular mitochondria with uniform size, intact mitochondrial cristae, intima and extima as well as complete nuclear membrane. The injury was obviously reduced in the APC+A/R group and SF+A/R group compared with the A/R group. The cellular structures in the Glib+APC+A/R group and Glib+SF+A/R group were similar to those in the A/R group. These findings showed that SF preconditioning is effective in protecting the myocardial cells of adult rats from A/R injury and the cardioprotective effect of SF preconditioning is related to the activation of intracellular K+ATP channel.
 

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