中国组织工程研究

中国组织工程研究 ›› 2015, Vol. 19 ›› Issue (5): 739-744.doi: 10.3969/j.issn.2095-4344.2015.05.015

• 器官移植动物模型 organ transplantation and animal model • 上一篇    下一篇

不同缺血后调适方案对大鼠模型肢体骨骼肌缺血再灌注损伤的影响

彭龙龙，阳富春，薄占东，谭  桢，姚  军，程建文，薛明强，赵劲民   

  1. 广西医科大学第一附属医院创伤骨科手外科，广西医科大学再生医学研究所，广西壮族自治区南宁市  530021
  • 修回日期:2014-12-10 出版日期:2015-01-30 发布日期:2015-03-02
  • 通讯作者: 阳富春，博士，教授，广西医科大学第一附属医院创伤骨科手外科，广西医科大学再生医学研究所，广西壮族自治区南宁市 530021
  • 作者简介:彭龙龙，男，1989年生，湖北省潜江市人，汉族，广西医科大学在读硕士，主要从事创伤修复与功能重建研究。
  • 基金资助:

    广西自然科学基金资助项目(2011GXNSFC018021)；国家自然科学基金资助项目(81260276)

Effect of different protocols of ischemic postconditioning on skeletal muscle ischemia/reperfusion injury in a rat model

Peng Long-long, Yang Fu-chun, Bo Zhan-dong, Tan Zhen, Yao Jun, Cheng Jian-wen, Xue Ming-qiang, Zhao Jin-min   

  1. Department of Trauma Orthopedics & Hand Surgery, The First Affiliated Hospital of Guangxi Medical University, Institute for Regenerative Medicine, Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China
  • Revised:2014-12-10 Online:2015-01-30 Published:2015-03-02
  • Contact: Yang Fu-chun, M.D., Professor, Department of Trauma Orthopedics & Hand Surgery, The First Affiliated Hospital of Guangxi Medical University, Institute for Regenerative Medicine, Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China
  • About author:Peng Long-long, Studying for master’s degree, Department of Trauma Orthopedics & Hand Surgery, The First Affiliated Hospital of Guangxi Medical University, Institute for Regenerative Medicine, Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China
  • Supported by:

    the Natural Science Foundation of Guangxi Zhuang Autonomous Region, No. 2011GXNSFC018021; the National Natural Science Foundation of China, No. 81260276

PDF

396

被引次数

4

摘要：

背景：近年来不少文献报道缺血后调适在多种组织器官包括心肌和骨骼肌都能激发自身内在的对缺血再灌注损伤的保护作用，而且不同方案引发的保护程度有差异，并表现出种属的特异性。
目的：比较不同缺血后调适方案对大鼠肢体骨骼肌缺血再灌注损伤的保护作用并优选最佳方案。
方法：健康成年雄性SD大鼠随机分为4组(n=9)：缺血再灌注组和不同缺血后调适3个组。各组大鼠均于右侧股动脉进行缺血再灌注实验操作，并分离左侧股动脉作为自身假手术对照，其中缺血再灌注组给予右侧股动脉缺血4 h再灌注24 h；缺血后调适3个组分别于4 h缺血后立即施加4个循环分别为10 s再灌/10 s缺血、30 s再灌/30 s缺血、1 min再灌/1 min缺血的后调适操作，再灌注24 h。各组实验结束后抽血检测乳酸脱氢酶，取样腓肠肌测算湿干质量(湿/干)比值、检测髓过氧化物酶和丙二醛，取样胫前肌电镜观察骨骼肌病理变化。
结果与结论：4个循环的30 s再灌/30 s缺血组湿/干值显著低于缺血再灌注组(P < 0.05)，其余2个缺血后调适组与缺血再灌注组差异不明显(P > 0.05)。血浆乳酸脱氢酶、组织髓过氧化物酶和丙二醛值缺血后调适3组均低于缺血再灌注组(P < 0.05)，缺血后调适组间未见明显差异。4个循环的30 s再灌/30 s缺血组电镜下骨骼肌线粒体嵴的空泡变性程度、肌原纤维结构清晰程度和细胞核完整性较缺血再灌注组均有明显改善，其余2个缺血后调适组超微结构较缺血再灌注组也有不同程度改善。结果提示缺血后调适对大鼠骨骼肌缺血再灌注损伤有保护作用，4个循环30 s再灌/30 s缺血方案的保护效果最明显，可作为进一步实验研究的基础。



中国组织工程研究杂志出版内容重点：肾移植；肝移植；移植；心脏移植；组织移植；皮肤移植；皮瓣移植；血管移植；器官移植；组织工程


全文链接：

关键词: 实验动物, 肌肉肌腱损伤模型, 缺血再灌注损伤, 骨骼肌, 缺血后调适, 方案优选, 大鼠, 国家自然科学基金

Abstract:

BACKGROUND: Recent studies have shown that ischemic postconditioning can trigger endogenous protection against ischemia/reperfusion injury in various organs including myocardium and skeletal muscle. However, there is variability in protective effects with different protocols and species specificity.
OBJECTIVE: To investigate effects of various ischemic postconditioning protocols on rat skeletal muscle ischemia/reperfusion injury and to select an optimal protocol.
METHODS: Healthy Adult male Sprague-Dawley rats were randomly allocated into four groups(n = 9): one ischemia/reperfusion injury group and three ischemic postconditioning groups. The rats in ischemia/reperfusion injury group were subjected to 4 hours of main ischemia in the right lower limbs via occlusion of the femoral artery with a clamp, followed by 24 hours of reperfusion, and sham surgery in the left lower limbs as self control. The surgical procedures in three ischemic postconditioning groups were same as in ischemia/reperfusion group except additional ischemic postconditioning interventions were applied at the onset of reperfusion after 4 hours of  ischemia with three different algorithms respectively: 4 cycles of 10 seconds reperfusion/10 seconds ischemia, 4 cycles of 30 seconds reperfusion/30 seconds ischemia, and 4 cycles of 1 minute reperfusion/1 minute ischemia. At the end of 24-hr reperfusion in each group, blood samples were taken from femoral vein for detecting serum lactate dehydrogenase(LDH), and tissue samples from gastrocnemius for measuring wet-dry weight (wet/dry)ratio, enzyme myeloperoxidase and malondialdehyde. Tibialis anterior muscle samples were also collected for observing the pathological changes in skeletal muscle under electron microscope. 
RESULTS AND CONCLUSION: The wet/dry ratio was significantly lower in the 30-second reperfusion/30-second ischemia group than in the ischemia/reperfusion injury group (P < 0.05). No significant difference was detected between other two ischemic postconditioning groups and ischemia/reperfusion group (P > 0.05). Plasma lactate dehydrogenase activity, gastrocnemius myeloperoxidase activity and malondialdehyde contents were lower in the three ischemic postconditioning groups compared to the ischemia/reperfusion injury group (P < 0.05). No significant difference was detectable among the three ischemic postconditioning groups. Vacuolar degeneration of skeletal muscle mitochondria, myofibrils structural clarity and integrity of the nucleus were apparently improved in the 30-second reperfusion/30-second ischemia group compared to the ischemia/reperfusion injury group under electron microscope. Ultrastructure changes of the other two ischemic postconditioning groups were improved to different degrees as compared with the ischemia/reperfusion injury group. The results indicate that ischemic postconditioning confers protection against skeletal muscle ischemia/reperfusion injury in the rat model. The protocol of four cycle of 30-second reperfusion/30-second ischemia is optimal, which can be used as a basis for further investigations.



中国组织工程研究杂志出版内容重点：肾移植；肝移植；移植；心脏移植；组织移植；皮肤移植；皮瓣移植；血管移植；器官移植；组织工程


全文链接：

Key words: Ischemia Reperfusion, Skeletal Muscle, Lactate Dehydrogenases, Myeloperoxidase, Malondialdehyde

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